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01 January 2011, Volume 15 Issue 1 Previous Issue    Next Issue

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Nucleofection effects on adult bone marrow-derived stem cells Li Zhen, Shen Xiao-tao, Cao Liang, Mao Ying-yu, Chen Si-yun, Zheng Xin, Cai Dong-qing 2011, 15 (1):  1-6.  Abstract ( 311 )   PDF (1757KB) ( 428 )   Save BACKGROUND: Adult bone marrow-derived stem cells are promising cell source in cell therapy. Tracking of adult bone marrow-derived stem cells is crucial to demonstrate the mechanism of stem cell migration and differentiation, and develop novel strategy for functional regeneration and stem cell therapy. OBJECTIVE: To explore effects of transfected adult bone marrow clonogenic stem cells (AMCSCs) on cell phenotype, proliferation and cardiac differentiation potential. METHODS: Plasmid-encoded reporter gene maxGFP was used for nucleofection of AMCSCs with U-23 program. Growth curves of AMCSCs before and after nucleofection were compared based on results of MTT assay. AMCSCs before and after nucleofection were treated with 3 μmol/L 5-azacytidine for inducing cardiac differentiation. The cardiac differentiation specific markers, GATA4 and MLC-2v, were applied to confirm cardiac differentiation by RT-PCR. The maxGFP transfected AMCSCs were conducted the intramyocardium injection into an adult Sprague-Dawley rat left anterior descending coronary artery (LAD) ligation model to trace the in vivo expression of transfected maxGFP gene. Fluorescence images of the injected heart were analyzed on days 2 and 7 postinjection. RESULTS AND CONCLUSION: At 24 hours following nucleofection, the transfection efficiencies of AMCSCs at passages 47 and 119 were 49.4% and 43.1%. On 5 hours, green fluorescence positive cells were observed. Following nucleofection, AMCSCs maintained round shape, and could adhere and show clone-shape growth. MTT assay results demonstrated that the passages 47 and 119 of AMCSCs exhibited similar growth curves before and after nucleofection. Mean population doubling time was 8.57 and 10.28 hours in passages 47 and 119 of AMCSCs prior to nucleofection, and 9.42 and 10.42 hours following nucleofection (P =0.551, P=0.774). RT-PCR results showed that AMCSCs expressed GATA4 before and after 5-azacitidine treatment prior to nucleofection, and strongly expressed MLC-2v strip after treatment. AMCSCs expressed GATA4 prior to and following 5-azacitidine treatment after nucleofection, and strongly expressed MLC-2v after treatment. No significant difference was determined in above-mentioned indexes prior to and following nucleofection. In vivo experiment results demonstrated that a few green fluorescence positive cells were apparent in injected myocardium on days 2 and 7 following transfected AMCSCs injection. Results indicated that nucleofection is an effective and fast method for transfection of exogenous DNA into cell. The AMCSCs which are experienced with nucleofection are able to maintain their morphology, proliferation and cardiac differentiation potential. However, only a few transfected AMCSCs express the transferred gene, GFP, after intramyocardium injection. Related Articles | Metrics

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Effects of injured spinal cord extracts on brain-derived neurotropic factor and myelin proteolipid protein in bone marrow mesenchymal stem cells Liu Ran, Fan Dong-yan, Jin Peng, Fan Hong-xue, Wang Ping 2011, 15 (1):  7-11.  Abstract ( 276 )   PDF (1393KB) ( 341 )   Save BACKGROUND: The component of injured spinal cord extracts is complex and includes some chemical substances or cytokines. It remains unclear if these factors may influence cellular proliferation and secretion of bone marrow mesenchymal stem cells (BMSCs) or not.   OBJECTIVE: To investigate the effect of injured spinal cord extracts on secretion of brain-derived neurotropic factor (BDNF) or myelin proteolipid protein (PLP) in BMSCs. METHODS: BMSCs were obtained from Wistar rats. After 3 generation of cell proliferation, BMSCs were induced with normal or injured spinal cord extracts for 20 days. Neuron specific enolase (NSE) positive cells were detected by immunofluorescence staining. Levels of BDNF and PLP in BMSCs cultured supernatant were evaluated using enzyme linked immunosorbent assay (ELISA) and BDNF and PLP mRNA was determined by real time polymerase chain reaction assay. RESULTS AND CONCLUSION: The rate of NSE positive cells and levels of BDNF and PLP that were treated with injured spinal cord extracts were greater significantly at different cultured time points than normal spinal cord extracts. These results showed that the injured spinal cord extracts could induce secretion of BDNF and PLP in BMSCs that is to the benefit of cellular differentiation into neuronal cells. Related Articles | Metrics

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Isolation and identification of rabbit bone marrow mesenchymal stem cells by low permeability combined with natural sedimentation Lin Chun-bo, Yang Yuan, Chen Wei-ping 2011, 15 (1):  12-16.  Abstract ( 276 )   PDF (1447KB) ( 603 )   Save BACKGROUND: The number of bone marrow mesenchymal stem cells (BMSCs) was few. 100 000 bone marrow karyocytes contain 1 BMSC. How to successfully isolate and identify BMSCs is a key in the study. OBJECTIVE: To establish a perfect culture system in vitro of BMSCs; BMSCs were identified from cell morphology, phenotype and functional aspects. METHODS: Low permeability principle was used to remove mixed cell, and combined with the natural sedimentation to isolate BMSCs. Their morphological characteristics were observed by light microscopy. The cell phenotype of BMSCs was determined by flow cytometry. BMSCs were induced into osteoblasts and adipocytes. Cell morphology was observed and alkaline phosphatase (AKP), Von Kossa, collagen typeⅠ, oil red O detection were performed. RESULTS AND CONCLUSION: BMSCs were identified as phenotype: CD29(+), CD45(-). Following being induced into osteoblasts, AKP, mineralized nodules and collagen type Ⅰ staining were detected positively; to adipocytes, the oil red O staining was detected positively. Results suggest that isolated BMSCs are non-hematopoietic stem cells with fibroblast-like cell morphology. These cells had the functions of differentiating into osteoblasts and adipocytes. Related Articles | Metrics

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Transdifferentiation of bone marrow adipocytes into osteoblasts Kang Peng-de, Pei Fu-xing, Yang Jing, Shen Bin, Zhou Zong-ke 2011, 15 (1):  17-22.  Abstract ( 349 )   PDF (1605KB) ( 609 )   Save BACKGROUND: Bone marrow adipocytes and osteoblasts are derived from bone marrow stromal cells. Both of them have homology and have transdifferentiation each other under a certain condition. OBJECTIVE: To investigate the activity of bone marrow adipocytes differentiation into osteoblasts and to explore the new treatment method for the steroid-induced osteonecrosis of the femoral head. METHODS: Osteogenic and adipogenic differentiation of confluent preadipocytes 3T3-L1 was performed. Changes in cell morphology were observed at various time points following culture. The levels of peroxisome proliferators activated receptor γ2 (PPARγ2), runt-related transcription factor 2 (Runx2), osteocalcin (OCN) and collagen type I (Col I) mRNA expression were investigated by RT-PCR at days 5 and 21 following culture. Western blotting was used to evaluate cellular PPARγ2, Runx2, OCN and Col I production on day 21 following osteogenic and adipogenic culture. Cultured cells received alkaline phosphatase staining, alizarin red staining and oil red O staining. 3T3-L1 osteogenic transdifferentiation and activity expression of osteoblasts were observed. RESULTS AND CONCLUSION: On day 5 following osteogenic induction, 3T3-L1 cells became spindle shape. Compared with control group, real-time quantity-PCR results demonstrated that PPARγ2 mRNA expression was reduced, but Runx2, OCN and Col I mRNA expression was slightly increased. On day 21, these changes were more significant. Western blot exhibited that Runx2, OCN and Col I expression was increased, but no PPARγ2 expression was detected. Positive expression of alkaline phosphatase staining was more. Alizarin red staining showed many scattered calcified nodes. Oil red O staining showed slight lipid droplet. Results indicated that mouse bone marrow stromal cells-derived preadipocytes 3T3-L1 can differentiate into osteoblasts with bioactivity to a certain degree following culture. There is the plasticity between adipocytes and osteoblasts. Related Articles | Metrics

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Angiogenesis in a rat model of focal cerebral ischemia following adipose tissue-derived neural stem cell transplantation Liu Bin, Liu Ning, Dong Jing, Wang Rui-min, Zhang Jin-xia 2011, 15 (1):  23-28.  Abstract ( 272 )   PDF (1623KB) ( 343 )   Save BACKGROUND: Adipose tissue-derived neural stem cells transplantation may improve neurological function in rats with cerebral ischemia injury, but its mechanism remains unclear. OBJECTIVE: To study the effects of human adipose tissue-derived neural stem cells transplantation on angiogenesis in rats after focal cerebral ischemia. METHODS: Adipose tissue-derived stromal cells cultured in vitro were differentiated into neural stem cells. A total of 60 healthy male Sprague-Dawley rats were assigned into four groups randomly: normal control group (n=6), sham-operated group (n=6), ischemia control group (n=24) and transplantation treated group (n=24). Models of 2-hour ischemia/reperfusion of rat middle cerebral artery were established by suture-occluded method in the ischemia control group and transplantation treated group. Ischemia control group and transplantation treated group were divided randomly into 7, 14, 21 and 28 days reperfusion groups (n=6). Middle cerebral artery was not occluded in the sham-operated group. On hour 24 following model induction, human adipose tissue-derived neural stem cell suspension (2×109 /L) was given in the transplantation treated group via the caudal vein. Saline was injected into the rats in ischemia control group via the caudal vein. Immunohistochemistry was used to identify microvessel density (MVD). The blood vessel proliferation in the rat focal cerebral ischemia area was observed.  RESULTS AND CONCLUSION: Immunohistochemistry results have exhibited that compared with the ischemia control group, the MVD was significantly greater at 7, 14, 21, 28 days after transplantation in the transplantation treated group (P < 0.05-0.01). Results indicated that neural stem cells differentiated from human adipose tissue-derived stromal cells transplantation can promote the angiogenesis in cerebral ischemia regions. Related Articles | Metrics

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Isolation and culture characteristics and influential factors of Sprague Dawley rat adipose tissue-derived mesenchymal stem cells  Hou Kai, Li Mei, Yang Yi-kun, Li Chun, Li Jin-ru, Huang De-qing 2011, 15 (1):  29-32.  doi: 10.3969/j.issn.1673-8225.2011. 01.006 Abstract ( 303 )   PDF (1228KB) ( 503 )   Save BACKGROUND: Adipose tissue-derived mesenchymal stem cells (AD-MSCs) could be obtained easily with abundant quantities; the fat cells left in the donation area have the capacity of proliferation and the injury is pretty mild. Therefore, AD-MSCs have been an ideal source of seed cells for tissue engineering. However, the isolation and culture of AD-MSCs still remain difficulties. OBJECTIVE: To explore the method for isolation and culture of AD-MSCs and observe their growing properties. METHODS: The adipose tissues were obtained from the inguinal groove, shoulder and abdominal cavity of Sprague Dawley rats aged 4 weeks and 10 months. Following washes and centrifugation, specimens were incubated in cell culture flask, and placed in 5%CO2 incubator. The liquid was replaced at 24, 48 and 72 hours. At a fusion of 80%, specimens were digested with 0.25% trypsogen-EDTA and passaged. Effects of age, gender, and the region of collecting adipose tissue, collagenase Ⅰ concentration and digestion time, and first time of liquid change on the culture of AD-MSCs were analyzed. RESULTS AND CONCLUSION: Primary AD-MSCs presented spindle or multangular shape. While AD-MSCs passaged and cultured longer, the newborn AD-MSCs, with polygon shape in clusters, joined to pieces and stretched into spindle or multangular shape gradually. A few fat cells appeared after 48 hours in culture and the cell number increased when the culture time was prolonged. However, about 90% of the isolated cells still kept the shape of AD-MSCs. The AD-MSCs could be isolated successfully from the fat harvested from the 4-week female Sprague Dawley rat abdominal cavity, and be passaged stably in vitro. The Sprague Dawley rats’ age, sex, collection site of adipose tissue, the concentration of collagenase Ⅰ and its digestion time, as well as the time of the first change of medium have an impact on the isolation and culture of AD-MSCs. Related Articles | Metrics

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Human umbilical cord mesenchymal stem cells transplantation improves lower limb ischemia of diabetic rabbits   Fan Ming-shi, Yang Xiao-feng, Wu Yan-xiang, Zhang Yi-bin, Xu Yi-feng, Song Nan, Huang Sheng-nan, Zhang Yue 2011, 15 (1):  33-36.  doi: 10.3969/j.issn.1673-8225.2011. 01.007 Abstract ( 368 )   PDF (1217KB) ( 625 )   Save BACKGROUND: The transplantation of bone marrow and autologous peripheral blood stem cells into the muscles of lower limbs of diabetes patients with lower limb ischemia can promote blood vessel regeneration, improve and recover the blood flow of affected limbs of diabetes patients with lower limb ischemia. However, the collection has great risk, and requires high levels in patients’ age, body condition and psychological reception degree. Compared with bone marrow and autologous peripheral blood stem cells, umbilical cord resource is abundant; cell collection is simple; immunogenicity is weak. OBJECTIVE: To investigate the feasibility that transplantation of human umbilical cord mesenchymal stem cells (HUCMSCs) for treatment of diabetic rabbit of lower limbs extremity arterial.  METHODS: Models of diabetic rabbits were established. Femoral arteries of lower limbs were ligated. Prepared HUCMSCs in sign of DIL were directly injected into the left hindlimb four-headed thigh muscles in diabetic rabbit models (cell quantity was 1.67×106 per point). Saline was directly injected into right lower limbs. At 2 and 4 weeks, adductor and gastrocnemius nuscle of the bilateral hindlimbs were collected. Angiogenesis of pathological sections was observed. Capillary density and skin temperature were measured using immunohistochemical staining for CD31. RESULTS AND CONCLUSION: Pathological sections exhibited that collateral vessels were significantly increased in the transplantation group compared with control group. Immunohistochemical staining for CD31 demonstrated that the capillary density was significantly higher in the transplantation group compared with control group. The skin temperature was significantly greater in the transplantation group compared with control group (P ≤ 0.05). Results suggested that HUCMSCs transplantation in the treatment of diabetic rabbit of lower limbs ischemia is an effective measure. Related Articles | Metrics

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Vascular endothelial cell growth factor combined with bone morphogenetic protein-2-modified bone marrow mesenchymal stem cells in repair of femoral head necrosis Cui Da-ping, Zhao De-wei 2011, 15 (1):  37-40.  doi: 10.3969/j.issn.1673-8225.2011. 01.008 Abstract ( 266 )   PDF (1065KB) ( 507 )   Save BACKGROUND: Vascular endothelial cell growth factor (VEGF) combined with bone morphogenetic protein (BMP) was used to repair necrotic bone, which can maintain the osteogenic phenotype of seed cells, and effectively secrete VEGF and BMP-2, and effectively promote blood vessel regeneration and contribute to formation and revascularization of tissue engineered bone tissues. OBJECTIVE: To observe the therapeutic effect on the treatment of avascular necrosis of the femoral head by using bone marrow mesenchymal stem cells (BMSCs) modified by VEGF-165 and BMP-2 in vitro. METHODS: The model of avascular necrosis of rabbit femoral head on right leg. There were single core decompression group, core decompression and BMSCs group, decompression with VEGF-165/BMP-2/BMSCs group. Necrotic bone was cleared out on arthroscope. Osteogenesis and angiogenesis were inspected by histology. RESULTS AND CONCLUSION: Arthroscope observation demonstrated that necrotic bone was cleared out in each group, and fresh blood was flowed out. Histomorphology determination showed that blood vessel number and new bone area in the repair region were significantly greater at various time points following transplantation in the decompression with VEGF-165/BMP-2/BMSCs group compared with single core decompression group and core decompression and BMSCs group (P < 0.05). These suggested that VEGF-165/BMP-2 gene transfection strengthened osteogenic effects of BMSCs, elevated number and quality of new bones and accelerated the repair of avascular necrosis of the femoral head. Related Articles | Metrics

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Bone marrow mesenchymal stem cells for the treatment of immune induced aplastic anemia Huang Ying, Zheng Cui-ling, Yang Shao-guang, Han Zhong-chao 2011, 15 (1):  41-45.  doi: 10.3969/j.issn.1673-8225.2011. 01.009 Abstract ( 281 )   PDF (1523KB) ( 452 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have immunomodulatory effects on lymphocytes and other immune-associated cells. Supportive effects of BMSCs on hematopoietic cells had been certified. Occurrence and development of aplastic anemia are closely associated with T cell hyperfunction-induced hematopoietic stem/progenitor cells. OBJECTIVE: To compare the therapeutic effects of BMSCs and whole bone marrow cells in immune-induced aplastic anemia mice. METHODS: Immune-induced aplastic anemia BALB/C mouse models were established. The BALB/C mice were randomly assigned to normal, control, BMSC injection and whole bone marrow cell groups. Therapeutic effects of BMSCs and whole bone marrow cells, volume of hemopoietic tissues, adipocytes and blood sinus structure were observed. RESULTS AND CONCLUSION: ①On day 20, all mice without treatment (8/8) and 4 of 8 mice which received BMSCs died, while 7 of 8 mice were infused with bone marrow cells and kept live. ②No significant difference in change of weight, blood cells, haematopoiesis was detected between BMSCs or bone marrow cells. ③In spleen cells of improved mice with BMSCs or bone marrow cells, the percentages of CD4+T cells, CD8+T cells and the ratio of CD4+/CD8+ were similar. In peripheral blood, the percentages of CD4+T cells and CD8+T cells were lower in BMSCs group than in normal group. The level of CD8+T cells was lowest in mice with bone marrow cells. These results indicate that BMSCs improve aplastic anemia symptom and recover hematopoiesis. The bone marrow cells were more effective than BMSCs in the therapeutical effect, but no significant difference was determined. Related Articles | Metrics

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Construction of rat models of myocardial ischemia/reperfusion injury following bone marrow mesenchymal stem cell pretransplantation for 1 week   Xu Feng, Wang Ai-ling, Chen Feng, Zhou Mei-ling 2011, 15 (1):  46-50.  doi: 10.3969/j.issn.1673-8225.2011. 01.010 Abstract ( 307 )   PDF (1597KB) ( 436 )   Save BACKGROUND: Blocked blood vessels were opened following acute myocardial infarction with thrombolysis or intervention method. It is difficult to reach a satisfactory outcome due to myocardial ischemia/reperfusion injury. OBJECTIVE: To observe the repair effect of bone marrow mesenchymal stem cells (BMSCs) transplantation on myocardial ischemia/reperfusion injury in rats. METHODS: BMSCs were cultured and expanded from healthy 3-week-old Sprague-Dawley rats in vitro. Recipient Sprague-Dawley rats were injected with BMSCs or serum-free DMEM. 1 week later, rat models of myocardial ischemia/reperfusion injury were established. Following myocardial ischemia for 0.5 hour and reperfusion for 2 hours, serum lactate dehydrogenase (LDH) activity, total superoxide dismutase (TSOD) vitality and malondialdehyde (MDA) levels of myocardial tissue were measured. Cell apoptotic index was examined by terminal deoxynucleotidyl transferase-mediated d-UTP nick end labeling. The expressions of Bcl-2 and Bax proteins were measured by immunohistochemical technique. RESULTS AND CONCLUSION: BMSCs not only had a high purification, homogeneity and stability, but also had unlimited proliferation. Compared with ischemia/reperfusion group, LDH leakage was reduced, TSOD activity was increased and MDA levels were diminished in the BMSCs group (P < 0.01). Apoptotic index was significantly lower in the BMSCs group compared with ischemia/reperfusion group (P < 0.01). Bcl-2 expressions were greater in the BMSCs group compared with ischemia/reperfusion group (P < 0.05), but the Bax/Bcl-2 ratio in the BMSCs group was lower than the ischemia/reperfusion group (P < 0.05). Results suggest that BMSCs transplantation can promote cell proliferation of rats with myocardial ischemia/reperfusion injury. Related Articles | Metrics

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Assessment for the reperfusion with sonoVue intravenous myocardial contrast echocardiography in patients with acute myocardial infarction undergoing intracoronary autologous bone marrow mononuclear cells transplantation: Compared with myocardial perfusion tomographic imaging  Qian Da-jun, Liu Huan, Zhou Da-qiong, Wu Peng-xi, Yang Zhen-yu, Wu Xiao-qing, Wang Qiang 2011, 15 (1):  51-54.  doi: 10.3969/j.issn.1673-8225.2011. 01.011 Abstract ( 266 )   PDF (1443KB) ( 344 )   Save BACKGROUND: Myocardial perfusion tomographic imaging and magnetic resonance imaging were commonly used to evaluate myocardial reperfusion following stem cell transplantation for treatment of acute myocardial infarction (AMI). However, the repetitiveness of this method is poor, and has radioactivity. OBJECTIVE: To estimate the reperfusion of intravenous myocardial contrast echocardiography (IMCE) in patients with AMI using intravenously infused SonoVue before and after intracoronary autologous bone marrow mononuclear cells transplantation. METHODS: IMCE was performed in 24 AMI patients before and after intracoronary autologous bone marrow mononuclear cell transplantation. The maximal amplitude score (A), the mean ascending slope of the curve (β) and the product of A×β were measured, and compared with results from myocardial perfusion tomographic imaging. Myocardial reperfusion was assessed. RESULTS AND CONCLUSION: IMCE results exhibited that A, β and A×β were significantly increased following transplantation (P < 0.01). The changes in β value were maximal following vessel unobstruction, and change range of A value was increased following 6-month follow-up. These indicated that perfusion speed was accelerated immediately following vessel unobstruction. Myocardial perfusion defect was obviously improved at 6 months following transplantation. Myocardial nuclide intake was enhanced immediately and at lag phase in the infarct region at 6 months following transplantation, and the score of single photon emission computed tomography imaging was significantly diminished (P < 0.05). These indicated that results of IMCE are basically identical to that of myocardial perfusion tomographic imaging, and IMCE can noninvasively assess microcirculation perfusion of transplanted myocardial cells. Related Articles | Metrics

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Allogeneic hemopoietic stem cell transplantation using mobilization with granulocyte colony stimulating factor and granulocyte-macrophage colony stimulating factor Liu Zhong-wen, Guo Jian-min, Yang Jing, Zhang Yin 2011, 15 (1):  55-58.  doi: 10.3969/j.issn.1673-8225.2011. 01.012 Abstract ( 295 )   PDF (614KB) ( 337 )   Save BACKGROUND: Currently, only granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) approved by Food and Drug Administration are used for the mobilization of peripheral blood hemopoietic stem cells and G-CSF alone or in combination with GM-CSF is the predominant mobilization regiments used in the allogeneic setting, but it might cause the donors some adverse events such as bone pain, muscular soreness and fever. OBJECTIVE: To retrospectively review the clinic results of allogeneic peripheral blood hemopoietic stem cell transplantation (allo-PBSCT) using mobilization with G-CSF and GM-CSF. METHODS: From January 2004 to October 2009, a total of 51 patients with hematological malignant diseases received allo-PBSCT from human leucocyte antigen-matched sibling donors mobilized with G-CSF and GM-CSF at the Department of Hematology, Henan Provincial People’s Hospital. We analyzed components in the allografts, hematopoietic reconstitution and the incidence of graft versus host diseases (GVHD). RESULTS AND CONCLUSION: After mobilizing 96 hours, the percentage of CD34+ cells in mononuclear cells was (0.97±0.13)% and the percentage of CD34+/CD38- cells in CD34+ cells was (37.49±4.03)%. Rapid hematopoietic reconstruction posttransplantation was negatively associated with infused total CD34+ and CD34+/CD38- cell number. Incidences of Grades Ⅰ, Ⅱ–Ⅳ acute GVHD were respectively 25.5% and 15.7% of patients. The incidence rates of limited and extensive chronic GVHD were 39.2% and 21.2% separately. These results suggest the combination regimen with GM-CSF and G-CSF appear to have a good effect in the mobilization of peripheral blood stem cells and be sufficient to ensure early hematopoietic reconstitution. The high doses of infused CD34+ and CD34+CD38- cells are likely beneficial to the prompt engraftment. Related Articles | Metrics

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Autologous stem cell orthotopic transplantation in treatment of young middle-age patients with ischemic stroke  Chen Yu-hua, Xu Li, Chen Qi-ming, Chen Shou-kang, Xu Wen-fang, Qu Hong-dang, Qian Wei-dong, Wei Dao-xiang, Liu Xiao-lin 2011, 15 (1):  59-62.  doi: 10.3969/j.issn.1673-8225.2011. 01.013 Abstract ( 274 )   PDF (680KB) ( 434 )   Save BACKGROUND: If endogenous stem cells levels were elevated in patients, and stem cells homing to a damaged site, and then self-repair ability was enhanced, which is accorded with reactive repair mechanism of self-damage, and can avoid immunologic rejection induced by allotransplantation. OBJECTIVE: To investigate safety and availability on autologous stem cell stimulated by granulocyte colony-stimulating factor (G-CSF) orthotopic transplantation in treatment of young middle-age acute ischemic stroke by observing functional recovery and infarct area changes. METHODS: A total of 40 cases of young middle-age acute cerebral infarction (middle cerebral artery territory) patients were divided into two groups randomly. In orthotopic transplantation group, 20 patients received subcutaneous G-CSF (15 μg/kg per day) injections for 5 days within 7-10 days following onset. The rest treatment was identical to the control group. In control group, patients were given conventional therapy of infarction. Before 10 days and 3 months after onset, National Institutes of Health Stroke Scale (NIHSS) and the Alberta stroke program CT score (ASPCTS) were performed. RESULTS AND CONCLUSION: No patients developed severe adverse reaction during the treatment. NIHSS of treatment group after 3 months was lower and ASPCTS was higher significantly than the control group (P < 0.05). Results indicate that it was safe and effective for young middle-age patients with acute cerebral infarction to be treated by autologous stem cells stimulated by granulocyte colony-stimulating factor orthotopic transplantation. The treatment can decrease infarct size and improve the recovery of neurological function. Related Articles | Metrics

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Comparison of hematopoietic stem cell transplantation and non-transplantation for treatment of severe aplastic anemia    Yan Hong-min, Liu Jing, Xue Mei, Wang Zhi-dong, Zhu Ling, Ding Li, Wang Heng-Xiang 2011, 15 (1):  63-67.  doi: 10.3969/j.issn.1673-8225.2011. 01.014 Abstract ( 434 )   PDF (523KB) ( 489 )   Save BACKGROUND: Hematopoietic stem cell (HSC) transplantation is an optimal therapeutic method of severe aplastic anemia. The transplantation of various sources-derived HSCs has been performed, including affinity haploid transplantation, unrelated transplantation. However, the reports on curative effects of HSCs combined with MSCs transplantation are individual case. OBJECTIVE: To retrospectively compare and analyze the curative effect of HSC transplantation and non-transplantation in treatment of severe aplastic anemia. METHODS: We studied 17 patients with severe aplastic anemia aged 3-53 years from April 2008 to April 2010. In them, 8 patients were subjected to HSC transplantation, and 9 patients were treated with non-transplantation. In the transplantation group, 4 patients received related HLA half-matched, 2 received HLA matched, and 2 received unrelated HSC transplantation. In all patients, 4 of them also received mesenchymal stem cell co-transplantation besides HSC infusion. The 9 patients in non-transplantation group received immunosuppressant and hematopoietic promotion treatment. RESULTS AND CONCLUSION: One 45-years old patient in the transplantation group had received non-transplantation for 11 months, but no effect was determined; thus, the patient received transplantation when this patient was combined with renal failure and lung fungous infection, and died of transplantation complication. The chromosome and DNA fingerprint detection of the remaining 7 patients indicated that complete donor implantation and rapid recovery of hematopoietic functions following HSC transplantation. The mean time of neutrophil exceeded 0.5×109/L and platelets recovery exceeded 20×109/L were 12 and 14 days for all transplantation patients, respectively after transplantation, and 11.6 days (neutrophil exceeded 0.5×109/L) and 11.6 days (platelets recovery exceeded 20×109/L) in HSC co-transplants with MSC respectively after transplantation. Four patients received degrees Ⅰ and Ⅱ acute graft versus host disease, and four received limited chronic graft versus host disease. The quality of life of post-transplant patients was good, without blood products infusion or severe infection or bleeding. The hematopoietic function of patients in the non-transplantation group was not recovered to normal. One patient died of cerebral hemorrhage and infection. The remaining patients had low quality of life, and should be hospitalized for symptomatic treatment, long-term discontinuous blood products infusion. Following treatment, many kinds of severe complications occurred. Results suggested that HSC transplantation is highly effective method for treating severe aplastic anemia. The hematopoietic function can be rapidly recovered. Graft versus host disease can be prevented and controlled, showing high quality of life. The curative effect is obviously better compared with non-transplantation. Related Articles | Metrics

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Influence of stromal cell-derived factor-1 alpha and its receptor CXCR4 on vascular smooth muscle cells and bone marrow stromal cells in common carotid artery balloon injury models   Cai Wen-wei, Fang Ning-yuan, Sheng Jing, Ma Shao-jun, Cheng Zhi-hui 2011, 15 (1):  68-73.  doi: 10.3969/j.issn.1673-8225.2011. 01.015 Abstract ( 288 )   PDF (2124KB) ( 377 )   Save BACKGROUND: Bone marrow stromal cells (BMSCs) can differentiate into endothelial progenitor cells by the action of cytokine and migrate to the injured site to participate in vascular intima repair. OBJECTIVE: To observe the influence of the interaction between stromal cell-derived factor-1α (SDF-1α) and its receptor CXCR4 on vascular smooth muscle cells and BMSCs in the rat common carotid artery balloon injury model. METHODS: Vascular smooth muscle cells and BMSCs were cultured in vitro according to different time points just after surgery (0), 1, 4, 7, 14 days and 1 month after balloon injury in rat common carotid artery. SDF-1α mRNA expressions were detected in vascular smooth muscle cells while CXCR4 mRNA expressions were detected in BMSCs. SDF-1α siRNA was transfected into vascular smooth muscle cells and transwell body was used to measure the chemotaxis of SDF-1α or vascular smooth muscle cells to BMSCs. RESULTS AND CONCLUSION: SDF-1α mRNA expression began to increase just after balloon injury, and then subsided gradually on day 7. CXCR4 mRNA began to express on day 4 after balloon injury and lasted. Transwell cabin demonstrated that SDF-1 chemotaxis increased in injured BMSCs than in normal ones; the antagonist of CXCR4 receptor AMD3100 could weaken SDF-1 chemotaxis significantly. The chemotaxis of injured vascular smooth muscle cells to BMSCs was stronger than SDF-1 (P < 0.05) and the action could be weakened by AMD3100 or transfection of SDF-1α siRNA in vascular smooth muscle cells (P < 0.05). Results indicated that SDF-1α/CXCR4 should be important in the process of BMSCs migration to injured vessels. Related Articles | Metrics

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Combination of basic fibroblast growth factor and nerve growth factor affects the differentiation of adult rat hippocampal neural stem cells into neuron-like cells Tong Lei, Ji Li-li, Xie Da-long, Gao Hai, Tong Xiao-jie 2011, 15 (1):  74-77.  doi: 10.3969/j.issn.1673-8225.2011. 01.016 Abstract ( 246 )   PDF (1158KB) ( 387 )   Save BACKGROUND: Differentiation of neural stem cells (NSCs) was mediated by many factors. Several factors could induce NSCs to differentiate into neurons in varying degrees and it is now a focus on the control of NSCs differentiation.   OBJECTIVE: To study the effects of combination of basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) on the differentiation of adult rat hippocampal NSCs into neurons. METHODS: The rat brain hippocampus was removed sterilely. Cell suspension was prepared and diluted when the diameter of the fourth passage of clone sphere was 200 μm by mixture of DMEM/F12 containing 2% B27, 20 µg/L epidermal growth factor (EGF) and 20 µg/L bFGF. Monoclonal cells were passaged. NSCs were divided into blank control, bFGF, NGF and bFGF+NGF groups. Immunocytochemistry for identification of NSCs was done to detect positive rate of neuron specific enolase (NSE) and the differentiation of NSCs into neurons.   RESULTS AND CONCLUSION: ①The monoclonal cells expressed nestin and the differentiated cells expressed NSE and glial fibrillary acidic protein. ②The proportions of NSE-positive cells in bFGF group, NGF group and bFGF+NGF group were much higher than in blank control group (P < 0.05), with the highest in bFGF+NGF group (P < 0.05). These indicate that bFGF and NGF can elevate the differentiation of NSCs into neurons, and their combination obtains better outcomes. Related Articles | Metrics

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Analysis of protein interaction networks related to core transcription factors target genes in embryonic stem cells Zuo Chang-qing, Wang Zong-gui, Wu Tie, Cui Liao 2011, 15 (1):  78-81.  doi: 10.3969/j.issn.1673-8225.2011. 01.017 Abstract ( 405 )   PDF (1136KB) ( 431 )   Save BACKGROUND: It is sufficient to reprogram somatic cells, through transduction of some core transcription factors, into pluripotent stem cells (iPS) that exhibit the essential characteristics of embryonic stem (ES) cells. At present, the complex mechanism is not yet fully understood. OBJECTIVE: To study the protein interaction networks related to core transcription factors target genes in embryonic stem cells and to obtain regulatory mechanism controlled “stemness”. METHODS: Non-redundant protein interaction data (NRPD) were obtained after removal of redundant data in BioGRID database. Protein interaction pairs, formed by the target genes, were extracted by perl program and the largest continuous protein interaction networks were obtained through searching NRPD using breadth-first search algorithm. At the same time, 1 000 random networks were analyzed and compared. At last, network visualization was analyzed through the Cytoscape software and network characteristics were explained using complex scale-free network model of Barabasi-Albert. RESULTS AND CONCLUSION: More protein interaction pairs and larger continuous protein network, statistically significant difference compared with random genes, were formed by core transcription factor target genes. The continuous protein network is scale-free characteristics of complex networks. This study has suggested that target genes may regulate synergistically “stemness” characteristics of embryonic stem cells through close interaction and forming a network module. Related Articles | Metrics

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Establishment of a mouse model of graft-versus-host disease Zhu Jie-lin, Zhang Peng, Chen Fu, Hou Ru-rong 2011, 15 (1):  82-86.  doi: 10.3969/j.issn.1673-8225.2011. 01.018 Abstract ( 358 )   PDF (1399KB) ( 663 )   Save BACKGROUND: Mouse graft-versus-host disease (GVHD) is the main complication of allogenic hematopoietic stem cell transplantation, presents the damages to multisystem (skin, esophago, stomach intestine, liver), and is one of reasons for death following allogenic hematopoietic stem cell transplantation. OBJECTIVE: To explore the establishment of the mouse model of GVHD, and to provide effective animal models for experimental GVHD research. METHODS: C57BL/6(H-2b)→BALB/C(H-2d) was selected as donor and recipient of complete allotransplantation. The GVHD models were established by injection of lymphocytes and bone marrow cells of the spleen of donor mouse. Whether GVHD models were established was assessed according to clinical situation and pathological examination. RESULTS AND CONCLUSION: The recipient mice which accepted above 5×106 donor spleen cells developed acute GVHD after transplantation. However, the time when clinical situation of GVHD appeared and life span were different. Results indicated that the transfusion of 5×106 spleen cells was adequate to establish the mouse model of GVHD. The mouse model is reliable to the experimental research of GVHD. Related Articles | Metrics

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Effects of siRNA target gene B7H1 on adhesion and migration of human placenta mesenchymal stem cells Zhang Si-ying, Wang Guo-yan, Du Hai-bo, Luan Xi-ying 2011, 15 (1):  87-91.  doi: 10.3969/j.issn.1673-8225.2011. 01.019 Abstract ( 502 )   PDF (1501KB) ( 265 )   Save BACKGROUND: Human placenta mesenchymal stem cells (hPMSCs) express costimulatory molecules B7H1 which plays an important role in its immunosuppressive capabilities. However, the effect of B7H1 on the biological behavior of hPMSCs has not been reported. OBJECTIVE: To investigate the effect of B7H1 blockade on adhesion and migration of hPMSCs. METHODS: hPMSCs were isolated from mature placenta by the method of digestion. Then in vitro hPMSCs were cultured, expanded and were used in test after third passage. Flow cytometry and RT-PCR analysis were used to investigate the expression of B7H1 on hPMSCs. Specific siRNAs of B7H1 were transfected into hPMSCs via cathodolyte liposome transfection method. RT-PCR was used to detect the changes of B7H1 gene expression. Then the adhesion of hPMSCs was determined by cell count, and the migration of hPMSCs was evaluated using Transwell method. RESULTS AND CONCLUSION: siRNA could effectively block the expression of B7H1 which highly expressed on hPMSCs. After seeded for half an hour, the cell adhesion rate had no significantly difference in the blockade group and the control group. For 1 hour or 3 hours, however, the adhesion rate of hPMSCs was significantly higher in the blockade group than in the control group (P < 0.05). Transwell assay indicated that the migration numbers of hPMSCs in the blockade group were significantly less compared with the control group under the culture conditions of SDF-1α, DMEM-LG complete medium or hPMSCs culture supernatant (P < 0.01). The results showed that the adhesion ability of hPMSCs was enhanced while the ability of migration was inhibited after B7H1 blockade and suggested that B7H1 plays an important role in the adhesion and migration of hPMSCs. Related Articles | Metrics

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Differentiation of mouse embryonic stem cells into cardiomyocytes in hanging drop culture system in vitro   Liu Guan-xin, Wang Fang-jie, Zhao Dan, Lu Qi 2011, 15 (1):  92-95.  doi: 10.3969/j.issn.1673-8225.2011. 01.020 Abstract ( 298 )   PDF (1216KB) ( 478 )   Save BACKGROUND: It is theoretical and clinical significance of studying the condition and technique of the differentiation of embryonic stem cells embryonic stem cell (ESC) into cardiomyocytes and to further elevate rate of differentiation and cell purity. OBJECTIVE: To set up a model of inducing ESC into cardiomyocytes and to provide a suitable method to study development of cadiomyocytes in vitro.  METHODS: Undifferentiated ESC were incubated in vitro by hanging drop-suspension-adherence method (hanging drop culture), then divided into six groups according to different inducers added in the culture system: group Ⅰ (salvia miltorrhiza linked with 5-azacytidine as inducer), group Ⅱ (salvia miltorrhiza and retinoic acid as inducer), group Ⅲ (5-azacytidine as inducer), group Ⅳ (salvia miltorrhiza as inducer), group Ⅴ (retinoic acid as inducer), and group Ⅵ (no inducer). After inducer addition, Cardiomyocyte marker proteins expression of α-actinin, myosin, and α-actin were detected by immunocytochemistry in the ninth day. RESULTS AND CONCLUSION: Spontaneously contracting cells appeared in thesecond day following inducer addition under inverted microscope and one embryoid body had one or more contraction foci. The immunocytochemistry showed the cardiomyocyte marker proteins expression of α-actinin, myosin, α-actin were positive. The rate of differentiation of group Ⅰ was highest, which showed significant difference compared with groups Ⅱ and Ⅵ (P  < 0.01), and exhibited significant difference compared with groups Ⅲ, Ⅳ and Ⅴ (P < 0.05). it suggested that traditional Chinese drug salvia miltorrhiza linked with 5-azacytidine as inducers in culture system of hanging drop cultures can improve rates of ESC into cardiomyocytes differentiation in vitro. Related Articles | Metrics

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Separation, culture and characteristics of epidermal stem cells in diabetes mellitus rats Zhu Fei-bin, Liu De-wu, Zhang Hong-yan, Peng Yan, Zhong Qing-ling, Li Yong-tie 2011, 15 (1):  96-99.  doi: 10.3969/j.issn.1673-8225.2011. 01.021 Abstract ( 355 )   PDF (1311KB) ( 398 )   Save BACKGROUND: Epidermal stem cells (ESCs) serve as skin tissues specific stem cells, it plays critical roles in wound repairing, but the study about separation and culture of diabetes mellitus skin-derived ESCs in vitro and biological characteristics is fewer.  OBJECTIVE: To explore the methods of separation and culture of diabetes mellitus rat ESCs, investigate basic biological characteristics, and provide the experimental evidences for diabetes mellitus mechanism research and non-healing wound treatment. METHODS: Sprague Dawley rats were randomly divided into diabetes mellitus group and normal control group. Diabetes mellitus rat model was made by intraperitoneal injection of streptozocin, and the normal control group was not treated. Full-thickness skins of the back of diabetes mellitus group rats and normal rats were taken respectively. Enzyme digestion combined with type IV collagen attachment method was used to separate, culture and choose rat ESCs. Morphologic change and cell colony formation were observed under inverted phase contrast microscope. Cell growth curve was detected by cell counting and cell colony formation rates were measured. The positive expressions of K19, β1-integrin were identified by immunocytochemical stain and picture analysis software. The integral absorbance value of positive cells was detected. RESULTS AND CONCLUSION: The number of primary anchorage-dependent ESCs in diabetes mellitus group was decreased, the colony forming efficiency of ESCs in diabetes mellitus group was significantly lower than that in the normal control group (P < 0.01). ESCs of both groups expressed K19 and β1-integrin. The integral absorbance values of positive cells for K19 and β1-integrin in ESCs of diabetes mellitus group were significantly lower than those in the normal control group (P < 0.01). Results indicate that fast separation and stable culture in vitro of diabetes mellitus ESCs could be achieved by enzyme digestion and type IV collagen attachment method. The multiplication capacity in vitro of rat ESCs in diabetes mellitus group was slower than normal skin and it may be one of important factors that diabetes mellitus wound is hard to heal. Related Articles | Metrics

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Effects of methylene blue photochemical virus inactivation on plasma components Jiao Hong-liang, Guan Fang-xia, Yang Bo, Li Jian-bin, Shan Hong, Du Ying, Hu Xiang 2011, 15 (1):  100-102.  doi: 10.3969/j.issn.1673-8225.2011. 01.022 Abstract ( 457 )   PDF (825KB) ( 385 )   Save BACKGROUND: Virus inactivation to blood is one of blood transfusion safety measures. The effect of virus inactivation on human plasma by methylene blue/light has been confirmed, but the report concerning virus inactivation to plasma on plasma components is very few. OBJECTIVE: To observe the effects of methylene blue photochemical virus inactivation to blood plasma on blood components’ structure and function. METHODS: A total of 40 blood samples were randomly selected. Fresh plasma from 400 mL whole blood within 6 hours was prepared and the quality was weighed for reserve samples, and then the filter with methylene blue virus inactivation was sterilely connected. The final concentration of methylene blue was 0.9-1.3 μmol/L. Plasma added to methylene blue virus inactivation into 4 ℃ was put on a shelf box, with beat frequency of 60 times/minutes. 32 000-38 000 Lx intensity of 4 ℃ visible light was used to irradiate for 35 minutes. The residual methylene blue and white blood cells were filtered out from the plasma after illumination by the virus inactivated filters, mixed and remained sample 10 mL, and immediately placed in -80 ℃ refrigerator. The amount of plasma samples before and after exposure, methylene blue concentration, F Ⅷ: C, F Ⅴ: C, VWF, Fib Content was detected. RESULTS AND CONCLUSION: After virus inactivated plasma, the recovery rate of plasma volume, F Ⅷ: C, F Ⅴ: C, VWF, Fib were (96.39 ± 1.73)%, (82.55 ± 9.25)%, (81.03 ± 15.27)%, ( 93.25 ± 6.17)%, (81.61 ± 14.25)%. The effect of methylene blue photochemical virus inactivation in plasma on most components of the plasma is not obvious, and meets the clinical demand for safe blood transfusion. Related Articles | Metrics

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Application of embryonic stem cells to reproductive medicine Liu Li-qing, Li Juan, Zhou Sheng-nian 2011, 15 (1):  113-115.  doi: 10.3969/j.issn.1673-8225.2011. 01.025 Abstract ( 248 )   PDF (512KB) ( 431 )   Save BACKGROUND: It has been reported that embryonic stem cells can differentiate into primordial germ cells in vitro culture and to further generate gametes then develop into blastocysts after fertilization, and produce live filial generation. OBJECTIVE: To expound the research progress of embryonic stem cells in reproductive medicine in recent years. METHODS: Tongfang database system was used to search for research articles published from 2000 to 2009 on germ cells and Chinese medicine related to the key words “germ cells, medicine”, and the language was limited to Chinese; the same searching PubMed database articles published from 1998 to 2009 with the key words of “embryonic stem cells, stem cells, germ cells, oocyte, sperm, differentiation”, and the language was limited to English. RESULTS AND CONCLUSION: Embryonic stem cells in vitro differentiation affected by internal and external factors, can be induced to differentiate into sperm and oocytes, is an ideal model on the study of early embryonic development in mammals. At present, it is feasible to in vitro culture primordial germ cells and gamete, but the characteristics and function of in vitro cultured germ cells remain unclear and it is necessary to further identify its safety and efficacy in clinic. Related Articles | Metrics

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Drug resistance and its mechanisms of cancer stem cells Bao Qiu-ping, Li Hui-min 2011, 15 (1):  116-119.  doi: 10.3969/j.issn.1673-8225.2011. 01.026 Abstract ( 448 )   PDF (538KB) ( 1265 )   Save BACKGROUND: Tumor stem cells not only associate with multidrug resistance in cancer, and have a complex multi-drug resistance OBJECTIVE: To review correlation of cancer stem cells to drug resistance and resistance mechanisms, and to provide new ideas for clinical therapy of tumor. METHODS: We searched PubMed Western database and FMJS database (2003-2010) with the key words of “cancer stem cell, multiple drug resistance, apoptosis, cell cycle”. Wanfang database (2000-2010) and CNKI database (2000-2010) were searched by computer with the key words of “tumor, cancer stem cells, multidrug resistance”. A total of 56 Chinese literatures, 372 foreign languages literatures were collected. Published earlier, repeated and similar studies were excluded and 29 literatures were included. RESULTS AND CONCLUSION: Stem cell research helps people deepen their knowledge of tumor occurrence, development and transfer. Tumor tissues have a small part of tumor cells that have self-renewal and unlimited proliferation ability as normal stem cells, but have a special mechanism of resistance. Tumor stem cells are closely related to tumor occurrence and treatment. However, complex multi-drug resistance of cancer stem cells requires further studies. Related Articles | Metrics

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Registered clinical studies of adult stem cells for non-malignant diseases Zhang Zhong-qian, Zhang Jun, Sun Hui, Jiang Susan, Zheng Yi-duan, Hu Sean 2011, 15 (1):  120-125.  doi: 10.3969/j.issn.1673-8225.2011. 01.027 Abstract ( 259 )   PDF (781KB) ( 407 )   Save BACKGROUND: Research on adult stem cells for the treatment of non-malignant disease grows quickly year-by-year. However, researchers are still working to identify potential indications for adult stem cells in non-malignant conditions OBJECTIVE: To provide consultative information on selecting potential indications of adult stem cells for non-malignant conditions. METHODS: On March 3rd, 2010, a search of the electronic database provided by the National Institute of Health website (http://www.clinicaltrials.gov) was conducted by using corresponding search formulas to define registered clinical research studies/trials in association with adult stem cells as an intervention in human subjects for non-malignant conditions. There was no restriction for language, research location, and research population race. RESULTS AND CONCLUSION: Of 1 540 studies identified in the initial search, 1 206 malignant conditions-related studies were excluded and a further 13 studies were excluded for lack of accordance with the searching purpose. A total of 321 research studies were entered into the analysis and reviewed. Among them, about a third of the trials were for cardiocerebrovascular diseases with the majority being ischemic cardiocerebrovascular diseases; the second third were for autoimmune disorders or graft versus host diseases after organ transplantation; the remaining third were for lower limb ischemic diseases (9%, 31/321), genetic disorders (9%, 31/321) and other diseases (22%, 73/321) of which the majority conditions were cirrhosis/hepatic failure, bone fusion operation, and type Ⅰ & Ⅱ diabetics. At present, clinical trials for adult stem cells in non-malignant diseases focus on the indications of cardiocerebrovascular diseases and peripheral vascular diseases. In addition, based on the theory of stem cells promoting tissue repair and regeneration, some of the studies were with a view to liver cirrhosis, bone fusion operation; and the idea of the immune downregulation of mesenchymal stem cells is still an important research concept. Related Articles | Metrics

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How long will the disease-specific induced pluripotent stem cells be applied in the clinical treatment of neurodegenerative diseases   Luo Hu, Liu Yu-kai, Li Ze-gui 2011, 15 (1):  126-129.  doi: 10.3969/j.issn.1673-8225.2011. 01.028 Abstract ( 367 )   PDF (571KB) ( 381 )   Save BACKGROUND: Both the medical treatments and the surgery can only relieve the symptoms but not prevent the progress of neurodegenerative diseases. Cell replacement therapy proceeds fast but its ethical problem and immune rejection also should be in consideration. However, the production of induced pluripotent stem cells provides us a new way to resolve these problems. OBJECTIVE: To review the basic situations and current treatment of neurodegenerative diseases and the possibility of the induced pluripotent stem (iPS) cells be used in regenerative medicine as a new cell resource. METHODS: The PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) and Wanfang database (http://www.wanfangdata.com.cn) were retrieved for articles published from 1995 to 2010 by computer. The English key words were “induced pluripotent stem cell, PD, ALS, SMA, degenerative disease”. The Chinese key words were “stem cells, Parkinson’s disease”. Totally 374 papers were primarily gotten. Finally, 29 papers were selected according to the inclusion criteria. RERULTS AND CONCLUSION: Patient-specific iPS cells can obtain the same genotype which is beneficial to the elucidation of the molecular mechanism of these diseases. These cells also posses the similar multi-directional differentiation ability as embryonic stem cells, which could be induced to dopamine neurons and relieve the clinical symptoms of Parkinson’s disease in animal model in vitro. The iPS cells have broad prospect in the treatment of neurodegenerative diseases. Related Articles | Metrics

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Bone marrow mesenchymal stem cells labeled with superparamagnetic iron oxide Hu Biao, Qiu Wei, Meng Zeng-dong 2011, 15 (1):  130-134.  doi: 10.3969/j.issn.1673-8225.2011. 01.029 Abstract ( 359 )   PDF (667KB) ( 688 )   Save BACKGROUND: At present, superparamagnetic iron oxide labeled bone marrow mesenchynal stem cells (BMSCs) have been rapidly applied in the treatment of heart, liver, kidney and central nervous system disease. OBJECTIVE: To summarize the present situation and progress about superparamagnetic iron oxide labeled BMSCs at home and abroad. METHODS: The database of CNKI and PubMed (2001-12/2009-12) was used to search the related articles about superparamagnetic iron oxide labeled BMSCs. Key words were “superparamagnetic iron oxide, stem cells, label, magnetic resonance imaging” that were searched in title and abstract. The articles related to superparamagnetic iron oxide labeled BMSCs published in recently five years, and for articles in the same field, those published recently or in authorized journals were selected. There were 243 articles after the initial survey. A total of 40 articles related to superparamagnetic iron oxide labeled BMSCs were included according to inclusion criteria for this review. RESULTS AND CONCLUSION: In previous studies addressing stem cell markers identified by light microscope, traditional histopathological examination was utilized, and the tissues were obtained in vivo. These are not suitable for dynamic observation, and not fit for clinical study. Superparamagnetic iron oxide can be used to effectively label living cells. This method cannot affect cell viability, growth, division, migration and differentiation, does not have cytotoxicity, with good safety, and magnetic resonance imaging can be utilized for in vivo tracer study. Related Articles | Metrics

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Bone marrow stromal stem cells and bone defect repair Li Pian, Rui Gang 2011, 15 (1):  135-138.  doi: 10.3969/j.issn.1673-8225.2011. 01.030 Abstract ( 337 )   PDF (562KB) ( 451 )   Save BACKGROUND: Bone marrow stromal stem cells can be widely obtained, easily separated, rapidly amplified, and with the potential of multiple directional differentiation, and are suitable for the characteristics of autotransplantation. Simultaneously, bone marrow stromal stem cells are suitable seed cells in bone tissue engineering. OBJECTIVE: To summarize and analyze the advantage of transgene or its complex stent for repair of bone defect using bone marrow stromal stem cells as seed cells. METHODS: Biomedical Bibliographic Database in Western Journals and China National Knowledge Infrastructure were searched for articles concerning isolation, culture and application of bone marrow stromal stem cells to bone defect as well as bone tissue engineering published from 1990 to 2009. The key words were “bone marrow stem cells, repairing, bone defect”. Repetitive articles were excluded. Finally, 28 articles were further summarized. RESULTS AND CONCLUSION: This article summarized the preponderance of bone marrow stromal stem cells as seed cells, isolation and culture of bone marrow stromal stem cells, osteogenic induction of bone marrow stromal stem cells and repair of bone defect using bone marrow stromal stem cells. The methods bring light prospects for the treatment of bone defect, such as bone marrow stromal stem cells as seed cells combined with suitable stent material, or bone marrow stromal stem cells as target cells to introduce endogenous target gene to induce osteogenic formation. However, some problems require further investigations, including purification of bone marrow stromal stem cells, proliferation and suitable condition for differentiation of bone marrow stromal stem cells, which kind of factors can effectively promote new bone formation, carrier material and in vivo implantation manner, as well as how to combine bone tissue engineering and gene therapy. Related Articles | Metrics

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Differentiation of mesenchymal stem cells into cardiomyocyte-like cells in vitro: Drug, microenvironment and method  Qin Jia-jia, Xian Shao-xiang, Huang Xi-wen, Sun Jing-he 2011, 15 (1):  139-142.  doi: 10.3969/j.issn.1673-8225.2011. 01.031 Abstract ( 282 )   PDF (662KB) ( 400 )   Save BACKGROUND: Cardiomyocyte cannot regenerate. Damaged cardiomyocytes cannot be repaired and differentiate. Clinical traditional Chinese medicine or western medicine, interventional therapy and surgery therapy cannot substitute necrotic myocardium. How to repair necrotic cardiomyocytes is an important topic in the cardiovascular research. Mesenchymal stem cells (MSCs) induced in vitro can differentiate into cardiomyocyte-like cells, which brings hope for the repair and regeneration of damaged cardiomyocytes. OBJECTIVE: To analyze the progress of inducing MSCs into cardiomyocyte-like cells in vitro to know where we should go in the future. METHODS: A computer-based online search of articles on the methods of inducing MSCs into cardiomyocytes in vitro was performed in CNKI and Elsevier-Sdol (2000-01/2010-08) with the key words of “mesenchymal stem cells, cardiomyocyte-like cells, induce, the progress of study”. A total of 185 articles on inducing MSCs into cardiomyocyte-like cells in vitro were collected, including 103 Chinese and 82 English. Outdate, repetitive and similar studies were excluded,and 32 were included. RESULTS AND CONCLUSION: Numerous studies have verified that MSCs could differentiate into cardiomyocytes in vitro under a certain condition. The induction method mainly contained differentiation inducer method and simulated cardiac microenvironment method. The efficiency of differentiation into cardiomyocytes should be elevated. The optimal induction drug and optimal in vitro microenvironment during the differentiation of MSCs into cardiomyocytes should be explored. Related Articles | Metrics

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Development of mechanical mechanisms of stem cells Bao Zhi-qiang, Ren Ming-ji, Li Jian-fu 2011, 15 (1):  143-146.  doi: 10.3969/j.issn.1673-8225.2011. 01.032 Abstract ( 215 )   PDF (542KB) ( 541 )   Save BACKGROUND: Recently, studies addressing molecular basis of various cell signal transduction and regulation mechanism of relevant functions have obtained important progress, but the biological behavior of stem cells is complicated. The related signal transduction pathway and mechanism of proliferation and differentiation remain unclear. OBJECTIVE: To review relevant literatures, and to summarize the present status and application prospect of stem cell related signal pathway affected by cytomechanics. METHODS: China National Knowledge Infrastructure and PubMed database were retrieved by computer for articles concerning mechanical effect of mechanisms for stem cells published from 2005 to 2010. The key words were “stem cell, signal transduction” or “stem cell, stress”. The articles on signal transduction pathway of stem cells published recently in the same field or published in the authorized journals were selected, and totally 179 articles were primarily selected. Finally, 33 articles were included in accordance with inclusion criteria. RESULTS AND CONCLUSION: Many different signal pathways had been involved in the proliferation and differentiation of stem cells. The stem cells could play an important role in the stem appreciation and differentiation which were modulated by several different signal pathways, including F-actin, Ca2+, MAPK, Wnt and Notch. The exact mechanism of mechanics signal transduction and gene expression under stress condition will be an important content in future stem cell research. Related Articles | Metrics

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Biological characteristics and differentiation potential of mesenchymal stem cells Ding Zhi, Yang Song-lin 2011, 15 (1):  147-150.  doi: 10.3969/j.issn.1673-8225.2011. 01.033 Abstract ( 299 )   PDF (596KB) ( 534 )   Save BACKGROUND: With the development of regenerative medicine and cell therapy, mesenchymal stem cells have become a hot spot because of its wide source, easy isolation, strong proliferation and multi-directional differentiation ability. OBJECTIVE: To overview the research progress in biological characteristics and differentiation potential of mesenchymal stem cells. METHODS: The databases of PubMed, Springer Link, and Wanfang were retrieved for papers concerning discovery, nomenclature, biological characteristics and differentiation potential of mesenchymal stem cells published from January 2000 to September 2010 with key words of “mesenchymal stem cells, differentiation, transdifferentiation”. A total of 132 articles were retrieved. RESULTS AND CONCLUSION: Mesenchymal stem cells have been found for a long period, and extensively distributed in vivo. Its proliferation ability can be affected by multiple factors. Mesenchymal stem cells can secrete various cytokines, produce a series of extracellular matrix molecule, and express many kinds of surface markers, but not unicity and belong to immune deficiency cells. The isolation and purification method included the whole bone marrow adherence screening method and density gradient centrifugation. Its identification should synthesize many aspects. With the exception of differentiation into mesodermal cell lineage, mesenchymal stem cells can transdifferentiate into neural cells, hepatocytes, islet cells and epithelial cells. Transdifferentiation mechanism mainly contains heterogeneous theory, nuclear reprogramming theory and embryonic stem cell residual theory. Related Articles | Metrics

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Neural stem cells for repairing spinal cord injury Li Wei, Jiang Qi-sheng 2011, 15 (1):  151-154.  doi: 10.3969/j.issn.1673-8225.2011. 01.034 Abstract ( 255 )   PDF (607KB) ( 396 )   Save BACKGROUND: The development of neurobiology and stem cell technique increases the number of spinal nerves, reduces glial scar and forms cavities by cell transplantation. OBJECTIVE: To review the identification and characteristics of neural stem cells (NSCs), the possible mechanism, clinical study and clinical application of NSCs for repairing spinal cord injury. METHODS: The key words were “neural stem cells, transplant, spinal cord injury”. The first author retrieved PubMed database and Wanfang database for articles of the identification and characteristics of NSCs, the possible mechanism, clinical study and clinical application of NSCs for repairing spinal cord injury published from 1997 to 2010. Articles published earlier, repetitive ones and similar studies were excluded. A total of 29 articles accorded with inclusion criteria were included for this review. RESULTS AND CONCLUSION: NSCs have the ability to differentiate into neurons, astrocytes, oligodendrocytes and can replace the damaged neural cells. The articles discussed the identification and characteristics of NSCs, the possible mechanism, clinical study and clinical application of NSCs for repairing spinal cord injury. Some problems should be solved in the future study, including long-term survival and phenotypic stability of stem cells-derived neurons or glial cells following transplantation, whether a few embryonic stem cells of escape differentiation and selective program will amplify in the transplant site and form tumors. Related Articles | Metrics

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Functional characteristics of promyelocytic leukemia protein isoforms  Zhao Liang, Zhang Xue-mei 2011, 15 (1):  155-158.  doi: 10.3969/j.issn.1673-8225.2011. 01.035 Abstract ( 337 )   PDF (487KB) ( 390 )   Save BACKGROUND: Each isomer of promyelocytic leukemia protein is various due to different structural domain sequence at C terminal, but the effects of each isomer remain unclear. OBJECTIVE: To retrospectively analyze the genetic transcription regulation, cell growth differentiation, cell apoptosis, immune reaction and its karyosome formation of promyelocytic leukemia protein isomer. METHODS: The first author retrieved PubMed Database and FMJS Database for articles concerning wildtype promyelocytic leukemia protein, promyelocytic leukemia protein karyoplast, function as well as structure, naming and function of promyelocytic leukemia protein isomer and their interaction published from 1992 to 2010. RESULTS AND CONCLUSION: Promyelocytic leukemia protein gene was expressed as potentially many alternatively spliced mRNAs, each of which encodes a distinct protein i.e. promyelocytic leukemia protein isoform. As each promyelocytic leukemia protein isoform shares an identical N-terminal region, which decides they have general character in certain function, such as cell apoptosis. However, all isoforms of promyelocytic leukemia protein differ from one another by C-terminal domain, which decides that each promyelocytic leukemia protein isoform has respective character in certain function, such as intrinsic antiviral activities, cell growth and differentiation. Further studies will be required to explore the concrete biochemistry mechanism of the function of varied isoforms. Related Articles | Metrics

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Combined transplantation of neural stem cells and olfactory ensheathing cells for the treatment of cerebral hypoplasia Tao Xiao-yu, Liu Bo, Teng Xiao-hua, Deng Shi-jie, Xie Miao-jun, Xu Rong, Lu Ming 2011, 15 (1):  159-162.  doi: 10.3969/j.issn.1673-8225.2011. 01.036 Abstract ( 341 )   PDF (603KB) ( 501 )   Save BACKGROUND: Recent years, stem cell replacement therapy furnishes a new method for neural regeneration. But until now, we have not found the reports about stem cell transplantation for treating cerebral hypoplasia. OBJECTIVE: To explore the effectiveness of combined transplantation of neural stem cells and olfactory ensheathing cells for the repair of cerebral hypoplasia. METHODS: In October, 2009, a cerebral hypoplasia patient was enrolled at the 163 Hospital of the Chinese People’s Liberation Army. We used the treatment of physical cooling, neurotrophic, maintenance of water-electrolyte balance, hyperbaric oxygen. We selected the point of puncture between the L3-4 of the spine and injected neural stem cells (about 8×107) and olfactory ensheathing cells (about 4×107) slowly into the subarachnoid space, once two weeks, twice as a course, totally one course. We assessed the results of transplantation treatment by drop of body temperature, fine motor, motor coordination and language expression. RESULTS AND CONCLUSION: The body temperature decreased to normal level, and muscular tension reduced obviously after one month, he could sit up by himself after two months, walk under the help of handrail, and have simple language expression after three months. These indicated that it is effective to treat cerebral hypoplasia and can recover central nerve function by combined transplantation of neural stem cells and olfactory ensheathing cells. Related Articles | Metrics

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Autologous transplantation of bone marrow mesenchymal cells-derived neural stem cells for treating seven patients with central nervous system diseases Cao Zhi-yong, Liu Han, Shan Jun-jun, Zhu Meng-hui, Wang Jun-ming, Chen Hao, Cheng Yan-bo, Fan Hong-bin, Dong Rui-guo, Geng De-qin 2011, 15 (1):  163-166.  doi: 10.3969/j.issn.1673-8225.2011. 01.037 Abstract ( 270 )   PDF (493KB) ( 532 )   Save BACKGROUND: Previous studies of animal models of nervous system diseases have shown that bone marrow mesenchymal cells-derived neural stem cells have a good effect on central nervous system diseases. OBJECTIVE: To observe the therapeutic effects of autologous transplantation of bone marrow mesenchymal cells-derived neural stem cells applied for central nervous system diseases. METHODS: Seven patients with central nervous system diseases were subjected to by autologous transplantation of bone marrow mesenehymal cells-derived neural stem cells from October 2008 to April 2009. Bone marrow puncture aspiration was used to collected 50-60 mL bone marrow blood, which was prepared into bone marrow mesenchymal stem cell suspension following isolation and culture in vitro. Neural stem cells were isolated and induced with self cerebrospinal fluid, and transplanted into patients by lumbar puncture via the subarachnoid cavity. Neurological impairment and side effects posttransplantation were observed prior to and following transplantation. RESULTS AND CONCLUSION: No adverse reactions were found in three patients after transplantation. The neurological function of the seven patients had been improved in 6-month follow-up. Results suggest that it is a safe, convenient and effective method to use autologous transplantation of bone marrow mesenchymal cells-derived neural stem cells to treat patients with neurological impairment induced by nervous system diseases. Related Articles | Metrics

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Application of umbilical cord mesenchymal stem cell transplantation in the treatment of two cases of hereditary spastic paraplegia  Yang Hua-qiang, Wang Yun-fu, Li Dong-sheng, Du Ling, Yuan Ya-hong, Jiang Hua 2011, 15 (1):  167-170.  doi: 10.3969/j.issn.1673-8225.2011. 01.038 Abstract ( 539 )   PDF (551KB) ( 769 )   Save BACKGROUND: Hereditary spastic paraplegia is a kind of nervous system genetic disease which has clinical and genetic heterogeneity. Now, the treatment effectiveness of this disease is poor. OBJECTIVE: To observe the clinical effect and safety of umbilical cord mesenchymal stem cells (UC-MSCs) transplantation in the treatment of hereditary spastic paraplegia (HSP). METHODS: Two patients with HSP received UC-MSCs transplantation which total cellular score of UC-MSCs was (2-6)×107 by intravenous infusion and lumbar puncture intrathecal injections. Both patients were followed up after transplantation. Clinical symptoms and various indexes were observed and literature review was performed. RESULTS AND CONCLUSION: After transplantation, the clinical symptoms of both patients improved obviously: muscle tonus of both lower extremities reduced obviously, independent ambulation independent of crutches and another person’s aid, and walking was stable. Various biochemical indicators were normal and both patients had no severe complications or clear side effects after transplantation. Both patients’ conditions were continuously catabatic and no recurrence was found after one year follow up. These indicated that UC-MSCs transplantation is effective and can ameliorate clinical manifestations and delay progression of the disease in the treatment of HSP. Related Articles | Metrics

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Granulocyte colony-stimulating factor and interleukin-3 secretion in bone marrow stromal cells following Danggui Buxue decoction-containing serum treatment Wang Tao, Chen Li, Wang Xiao-ling, Feng Li, Chen Yu-hao, Tian Chen 2011, 15 (1):  171-173.  doi: 10.3969/j.issn.1673-8225.2011.01.039 Abstract ( 371 )   PDF (262KB) ( 379 )   Save BACKGROUND: Experiment suggests that the active component of Danggui Buxue decoction, i.e. polysaccharides, can significantly promote proliferation and differentiation of hematopoietic stem cells and hematopoietic progenitor cells, and regulate hematopoietic microenvironment. OBJECTIVE: To investigate the effect of serum containing Danggui Buxue decoction on granulocyte colony-stimulating factor (G-CSF) and interleukin-3 (IL-3) secretion in mice bone marrow stromal cells (BMSCs). METHODS: BMSCs were isolated and cultured in 96-well culture plate. BMSCs morphology and growth were observed by phase contrast microscopy. The cells were divided into four groups and cultured with different concentrations of the serum containing Danggui Buxue decoction. MTT method was used to observe the proliferation of BMSCs, and ELISA method was used to observe the expression level of G-CSF and IL-3. RESULTS AND CONCLUSION: Serum containing Danggui Buxue decoction significantly promoted BMSCs proliferation, and increased the expression level of G-CSF and IL-3 secreted from BMSCs in a dose-dependent manner. Notably, serum containing equivalent dose drugs significantly promoted BMSCs proliferation, and G-CSF and IL-3 expression compared with other groups. Higher concentration weakens cell proliferation and secretion effects. Related Articles | Metrics

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In vitro amplification and biological characterization of rabbit corneal limbal epithelial stem cells Mo Lian-jie, Ye Yu-feng, Ke Li-qin, Ren Wang-fang, Zhang Chun-fang, Wu Lian-bao,Zhang Fang-hua, Liu Xiao-ling 2011, 15 (1):  174-178.  doi: 10.3969/j.issn.1673-8225.2011.01.040 Abstract ( 396 )   PDF (662KB) ( 358 )   Save BACKGROUND: How to establish a stable in vitro culture system, including location of corneal limbal epithelial stem cells, in vitro sample harvest, in vitro culture, vector selection, as well as identification methods, play a key role in corneal limbal epithelial stem cells culture. OBJECTIVE: To culture the isolated rabbit corneal limbal epithelial stem cells and to identify the biological properties of cultured cells. METHODS: The primary rabbit cornel limbal epithelial stem cells were isolated and cultured with tissue inoculation using human amniotic membrane as vector. The growth features of cells were observed under an inverted microscope. The morphology of cells was observed by hematoxylin-eosin staining and a scanning electron microscope. Furthermore, the monoclonal antibody AE5 and P63 two-step immunohistochemical staining were used to identify limbal epithelial stem cell protein expression. RESULTS AND CONCLUSION: The rabbit corneal limbal epithelial stem cells could be successfully cultured and maintained a relatively high value-added potential in vitro. Rabbit corneal limbal epithelial stem cells cultured on the amniotic membrane pull netted cellular layer. The AE5 monoclonal antibody positive rate of primary cultured cells was about 5% and P63 monoclonal antibody positive up to 90%. AE5-positive rate increased and P63-positive rate decreased with the increase in the number of subculture. The rabbit limbal epithelial stem cells can be successful culture and amplified on human amniotic membrane in vitro by limbal tissue culture method. The cultured cells maintain the characteristics of corneal epithelial cells. The rabbit corneal limbal epithelial stem cells can form grafts on the amniotic membrane. Related Articles | Metrics

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Effects of mycobacterium phlei on proliferation of dendritic cells derived from human umbilical cord blood in vitro Wang Ling-zhen, Wang Gui-yun, Pang Xiu-ying, Zhao Yan-xia, Sun Li-rong 2011, 15 (1):  179-182.  doi: 10.3969/j.issn.1673-8225.2011.01.041 Abstract ( 332 )   PDF (455KB) ( 367 )   Save BACKGROUND: The central position of dendritic cells (DCs) has aroused increasing attention due to strong antigen presentation capability in anti-tumor. However, how to obtain enough functional DCs and reports regarding immunomodulator with low toxicity are few.   OBJECTIVE: To investigate the effect of mycobacterium phlei F.U.36 (Utilins) on the proliferation and maturity of DCs derived from human umbilical cord blood in vitro. METHODS: The mononuclear cells were isolated from human umbilical cord blood using Ficoll-Hypaque method and cultured with Utilins, cytokine (human recombinant granulocyte/macrophage colonystimulating factor + recombinant human tumor necrosis factor-α + recombinant human interleukin-4), or combination cytokine + Utilins, respectively. In addition, cells cultured with RPMI-1640 served as controls. Morphological features and growth of DCs were observed by an inverted microscope. The phenotype changes of DCs, such CD1a and HLA-DR, were detected by flow cytometry at 9 days after culture. Moreover, DCs were stained by Wright-Giemsa and observed under oil lens. RESULTS AND CONCLUSION: Typical DCs with high expressions of CD1a and HLA-DR were obviously detected in all experimental groups except the control group. The positive rates of CD1a and HLA-DR in the Utilins group were higher than those in the control group, but lower than the cytokine group (P < 0.05). The HLA-DR positive rate of the combination group was higher than that of the cytokine group (P < 0.05). The results revealed that, Utilins can not only promote the proliferation of DCs derived from human umbilical cord blood in vitro but also cooperate with cytokines to induce the maturity of DCs. Related Articles | Metrics

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Effects of CpG oligonucleotides on the function of peripheral blood mononuclear cells in patients with type 1 diabetes mellitus versus healthy volunteers Tian De-zeng, Liu Ming-zhe, Zhang Yong-xuan, Wei Xiao-hua, Ren Bao-xian, Wang Hai-min 2011, 15 (1):  183-186.  doi: 10.3969/j.issn.1673-8225.2011.01.042 Abstract ( 247 )   PDF (740KB) ( 317 )   Save BACKGROUND: CpG oligonucleotide has been shown to strengthen the function of peripheral blood mononuclear cells (PBMCs), but its effects on type 1 diabetes mellitus has been rarely reported. OBJECTIVE: To investigate the effects of CpG oligonucleotide on the expression of interferon γ (IFN-γ), interleukin (IL) -12 and IL-10 in PBMCs in patients with type 1 diabetes mellitus versus healthy controls. METHODS: PBMCs were isolated from patients with type 1 diabetes mellitus and healthy controls and then cultured in RPMI-1640 with non-stimulator (control group) and CpG oligonucleotide (CpG oligonucleotide group), respectively. The mRNA expression of IFN-γ, IL-10, and IL-12 in PBMCs was detected by reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: mRNA expression of IFN-γ and IL-10 was significantly lower in patients with type 1 diametes mellitus than in healthy controls (P < 0.01). In the CpG oligonucleotide group, the mRNA expression of IFN-γand IL-12 was significantly higher than in the healthy control group (P < 0.01), but the mRNA expression of IL-10 was similar to that in the healthy control group (P > 0.05). These findings demonstrated that CpG oligonucleotide can promote the production of IFN-γ and IL-12 in PBMCs of type 1 diametes mellitus. Related Articles | Metrics

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Surface marker changes in human umbilical cord-derived mesenchymal stem cells after cryopreservation and resuscitation 2011, 15 (1):  187-190.  doi: 10.3969/j.issn.1673-8225.2011.01.043 Abstract ( 366 )   PDF (579KB) ( 397 )   Save BACKGROUND: Mesenchymal stem cells are the stem cells that possess the capability for self-renewal and multi-directional differentiation. Umbilical cord is the tissue outside the embryos and would be fallen off after parturition. In addition, it has wide source and no ethical restriction, so it is promising to be the first choice for mesenchymal stem cells.  OBJECTIVE: To detect the surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) prior to and after cryopreservation and resuscitation. METHODS: After isolation and culture, morphology of the primary, P4 and P8 hUCMSCs was observed prior to cryopreservation and after resuscitation. Surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of primary, P4, and P8 hUCMSCs were detected through the use of flow cytometry prior to cryopreservation and after resuscitation RESULTS AND CONCLUSION: hUCMSCs prior to cryopreservation and hUCMSCs of different passages after resuscitation present the same phenotype, i.e., positive for CD29, CD44, CD49e, CD73, and CD90, and negative for CD34, CD45, CD271. These findings suggest that primary hUCMSCs do not present changes in surface markers after cryopreservation and resuscitation. Related Articles | Metrics