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08 May 2017, Volume 21 Issue 13 Previous Issue    Next Issue

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Effect of continuous versus intermittent force on the proliferation and differentiation of bone marrow stromal stem cells Chen Jun-liang, He Yun, Xiao Jin-gang 2017, 21 (13):  1969-1973.  doi: 10.3969/j.issn.2095-4344.2017.13.001 Abstract ( 298 )   PDF (958KB) ( 257 )   Save BACKGROUND: In the process of distraction osteogenisis, continuous and intermittent force can be used. However, which one is more beneficial for bone formation is still controversial. OBJECTIVE: To investigate and compare the effect of continuous and intermittent force on the proliferation and differentiation of bone marrow stromal stem cells (BMSCs). METHODS: Rabbit BMSCs were obtained using density gradient centrifugation method. The passage 3 BMSCs were seeded onto 6-well cell culture plates for mechanical investigation. Then, the cells were randomly divided into three groups. Group A was a control group without any treatments. Group B was the continuous force group with a centrifugal force (1 100 r/min, relative centrifugal force 20×g) applied 4 hours per day. Group C was the intermittent group with an centrifugal force (1 100 r/min, relative centrifugal force 100×g) applied twice in 4 hours and lasting 24 minutes once. After 3 days of culture under mechanical force, the proliferation of BMSCs and the expression of alkaline phosphatase and osteocalcin were detected and compared. RESULTS AND CONCLUSION: The centrifugal force promoted the ostegenic differentiation and proliferation of BMSCs and the expression of alkaline phosphatase significantly increased compared with the control group (P < 0.05). Compared with the intermittent force, the continuous force showed better effect on osteogenic differentiation and proliferation of BMSCs as well as the activity of alkaline phosphatase (P < 0.05).   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Cardiomyocyte-like differentiation of bone marrow mesenchymal stem cells:an association between related gene expression and cell morphological changes Xu Yi-chen, Liu Ling-long, Zhao Wen-jing, Wang Hui-feng, Wei Ya-shu, Chen Wei-ping 2017, 21 (13):  1974-1979.  doi: 10.3969/j.issn.2095-4344.2017.13.002 Abstract ( 332 )   PDF (1217KB) ( 204 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have been induced to differentiate into cardiomyocyte-like cells in vitro. OBJECTIVE: To explore the association between GATA-4, Nkx-2.5 and α-myosin heavy chain (α-MHC) expression and cell morphological changes and structure formation in the process of BMSCs differentiation into cardiomyocyte-like cells. METHODS: By using myocardial lysate, BMSCs were induced to differentiate into cardiomyocytes. Immunocytochemistry staining was used to detect cardiac troponin T (cTnT) and connexin43, for the identification of cardiomyocytes. In the process of directional differentiation, RT-PCR was used to detect the expression of GATA-4, Nkx2.5 and α-MHC. RESULTS AND CONCLUSION: During the directional differentiation of BMSCs, the cells were changed from long fusiform to short rod, forming protrusions that were interconnected to form mesh-like, bamboo-like or myotube-like structure. When the cells were interconnected like a bamboo, cTNT and connexin43 positive cells were visible, and then the number of positive cells increased with the presence of myotube-like structure. RT-PCR results showed that during the induced directional differentiation of BMSCs, GATA-4, Nkx2.5 and α-MHC mRNA levels increased continuously. When interconnected cells formed a mesh-like structure, GATA-4 expression reached the peak and then kept a high level. When adjacent cells were fused into a myotube-like structure, α-MHC reached the peak. Additionally, the expression of Nkx2.5 presented a time-dependent increase trend. Overall, during the induced differentiation of BMSCs into cardiomyocyte-like cells, the expression of cardiomyocyte specific genes, characterized by temporality and spatiality, is related to the changes of cell morphology and special structure formation.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Calcitonin gene-related peptide, a sensory neurotransmitter, promotes proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells from ovariectomized rats Chen Jie, Ding Yu-xiang, Liu Yan-pu, Ma Ge, Liu Wei 2017, 21 (13):  1980-1985.  doi: 10.3969/j.issn.2095-4344.2017.13.003 Abstract ( 289 )   PDF (4613KB) ( 202 )   Save BACKGROUND: Calcitonin gene-related peptide can promote the biological activity of osteoblasts. Here, we attempt to further study the effect of calcitonin gene-related peptide on osteogenic differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To observe the effects of exogenous calcitonin gene-related peptide on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells from ovariectomized rats. METHODS: An ovariectomized osteoporosis model in female Sprague-Dawley rats was employed, which was testified by micro-CT. Bone marrow mesenchymal stem cells were obtained from the femur of rats undergoing sham operation and ovariectomy, and then cultured and induced in the culture medium and osteogenic medium containing different concentrations of calcitonin gene-related peptide, respectively. The cell proliferation was detected using MTT method. After 7 days of osteogenic induction, real-time RT-PCR and western blot were used to detect Runx2 and Osterix expression at mRNA and protein levels. After 21 days of osteogenic induction, the osteogenic differentiation potentials of the cells were detected using alizarin red staining. RESULTS AND CONCLUSION: The osteoporosis model was successfully established by bilateral ovariectomy in the female rats. The proliferative and osteogenic differentiation potentials of bone marrow mesenchymal stem cells were dose-dependently enhanced by the different concentrations of calcitonin gene-related peptide. To conclude, the calcitonin gene-related peptide can increase bone formation by promoting the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells directly.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of bone marrow mesenchymal stem cells transfected by Zrt/Irt-like protein 1 on adipogenic differentiation via tumor necrosis factor alpha signaling pathway Zhang Xiao-yun, Li Jin-huan, Yuan Zhen-zhong, Chen Yue-ping, Xia Tian, Zhuo Ying-hong, Feng Yang, Lan Jiao, Dong Pan-feng, Zhao Bin 2017, 21 (13):  1986-1991.  doi: 10.3969/j.issn.2095-4344.2017.13.004 Abstract ( 346 )   PDF (5065KB) ( 195 )   Save BACKGROUND: Previous studies have confirmed that ethanol can promote adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and up-regulate the expression of PPARγ and aP2 in the tumor necrosis factor α (TNF-α) signaling pathway. As a member of the ZIP protein family, Zrt/Irt-like protein 1 (ZIP1) is closely related to bone metabolism and osteogenic differentiation. OBJECTIVE: To study the effect of BMSCs transfected by ZIP1 on TNF-α signaling pathway in the process of adipogenic differentiation. METHODS: The BMSCs from rabbits were isolated and cultured under different concentrations of alcohol (0.03, 0.09, 0.15, 0.21 mol/L), followed by transfection by ZIP1 siRNA and ZIP1 expression vector. RESULTS AND CONCLUSION: After culture in alcohol, the expression levels of aP2 and PPARγ proteins were both significantly increased (P < 0.05), and the level of triglyceride was increased in all alcohol groups except for 0.03 mol/L alcohol group (P < 0.05). After siRNA transfection, the expression levels of aP2 and PPARγ as well as the level of triglyceride were increased significantly in all the alcohol groups (P < 0.05); however, ZIP1 transfection decreased the expression levels of aP2 and PPARγ proteins (P < 0.05). To conclude, ZIP1 siRNA could promote the adipogenic differentiation of BMSCs through the activation of TNF-α signaling pathway.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Vitamin C treatment promotes cell proliferation of human adipose-derived stem cells under high glucose conditions Li Jiang-feng, Ding Shi-chao, Qi Ya-wei, Li Jin, Zeng Guo-fang, Lai Qiao, Liu Li, Zhang Pei-hua 2017, 21 (13):  1992-1997.  doi: 10.3969/j.issn.2095-4344.2017.13.005 Abstract ( 282 )   PDF (4493KB) ( 291 )   Save BACKGROUND: Although vitamin C has an anti-oxidation role and can promote cell proliferation, there is a lack of research about the promoting effect of vitamin C on the proliferation of adipose-derived stem cells under high glucose conditions and the related molecular mechanisms. OBJECTIVE: To explore the promoting effect of vitamin C on the proliferation adipose-derived stem cells treated by the high glucose and the related molecular mechanisms.  METHODS: Passage 3 human adipose-derived stem cells were cultured under high glucose conditions and then treated with different concentrations of vitamin C (0, 100, 150, 200, 250, 300 μmol/L). Cells cultured under low glucose conditions acted as controls. The expression levels of p-ERK and p-AKT proteins were detected by western blot. MTT method was used to choose the optimal concentration and time of vitamin C for all the subsequent tests. Human adipose-derived stem cells cultured under high glucose conditions were divided into four groups, and cells in blank control group had no treatment. Cells in the other three groups were treated with the optimal concentration of vitamin C (vitamin C group), LY294002+the optimal concentration of vitamin C (LY294002 group), or U0126+the optimal concentration of vitamin C (U0126 group) for 48 hours. EdU staining assay was used to detect the cell proliferation of human adipose-derived stem cells.  RESULTS AND CONCLUSION: (1) Cell counting kit detection: We found that high glucose reduced the proliferation of human adipose-derived stem cells, and vitamin C promoted the proliferation of these cells. The best concentration of vitamin C was 200 μmol/L and the optimal effect time was 48 hours. (2) Western blot detection: Compared with the 0 μmol/L vitamin C group, the level of p-ERK in the 200 μmol/L vitamin C group was upregulated significantly (P < 0.01), while no significant expression change in p-AKT protein was found in control, 0 and 200 μmol/L vitamin C groups. (3) EdU test: the number of EdU positive cells was significantly higher in the vitamin C, LY294002, and control groups compared with the blank control group (P < 0.01). Moreover, compared with the vitamin C group, the EdU positive cells in the U0126 group were decreased significantly in number (P < 0.01). In conclusion, the ERK/MAPK signaling pathway is involved in the promotion effect of vitamin C on the proliferation of human adipose-derived stem cells under high glucose conditions.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Autologous CD34+ cell transplantation promotes angiogenesis in older adult patients with atherosclerotic ischemia: study protocol for a prospective, single-center, open-label, randomized controlled clinical trial Zhou Chen-hong, Xu Li-li, Hao Xiu-xian, Sun Xiao-juan, Guo Ming-jin, Liu Bing 2017, 21 (13):  1998-2002.  doi: 10.3969/j.issn.2095-4344.2017.13.006 Abstract ( 270 )   PDF (910KB) ( 289 )   Save BACKGROUND: The main clinical manifestation of senile arteriosclerosis obliterans is lower limb ischemia, which is currently difficult to treat. One method is by autologous stem cell transplantation into the muscles of ischemic limbs to improve the formation of new capillaries and restore lower limb blood flow. Endothelial progenitor cell marker CD34+ cell transplantation has been shown to promote angiogenesis in ischemic limbs. Therefore, we propose that peripheral blood autologous CD34+ cell transplantation in older adult patients with atherosclerotic ischemia could effectively promote angiogenesis. OBJECTIVE: To assume that peripheral blood autologous CD34+ cell transplantation in the elderly with atherosclerotic ischemia could effectively promote angiogenesis. METHODS: This is a prospective, single-center, open-label, randomized, and controlled clinical trial that will be completed at the Qingdao No. 9 People’s Hospital, China. Twenty older adult patients with atherosclerotic lower limb ischemia will be randomized into two groups. In the cell transplantation group (n=10), peripheral blood CD34+ cells transfected with vascular endothelial growth factor 165 (VEGF165) gene will be intramuscularly transplanted into the ischemic limbs in older adult patients with atherosclerotic lower limb ischemia. In the control group (n=10), normal saline will be intramuscularly injected into the ischemic limbs. All patients will be followed up for 6 months. The primary outcome will be ankle-brachial indices before and 6 months after transplantation to assess lower limb ischemia in both groups. The secondary outcomes will be the number of microvessels in the lower limb muscles before and 6 months after transplantation, the morphology of new blood vessels revealed by CT angiography, the number of VEGF-immunoreactive cells 6 months after transplantation and the incidence of adverse reactions. The trial was registered at the ClinicalTrials.gov (identifier: NCT03098771), and the study protocol was approved by the Ethics Committee of Qingdao No. 9 People’s Hospital of China. All protocols will be in accordance with Declaration of Helsinki, formulated by the World Medical Association. All patients will be informed of study protocols and provide a written informed consent prior to the beginning of the trial. DISCUSSION: This trial will begin in January 2018 and finish in December 2019. We aim to quantify the effects of VEGF165 gene-modified CD34+ cell transplantation in the treatment of older adult patients with atherosclerotic ischemia to develop a new effective treatment of lower limb ischemia.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Psoralen reverses glutathione-S-transferase π-mediated multidrug resistance in breast cancer stem cells Hua Yi-tong, Wang Xiao-hong, Xu Cheng-feng, Cheng Kai, Sun Hong-guang, Zhang Ying-zhe, Liu Jian, Yang Zhen-lin 2017, 21 (13):  2003-2008.  doi: 10.3969/j.issn.2095-4344.2017.13.007 Abstract ( 247 )   PDF (1145KB) ( 330 )   Save BACKGROUND: Breast cancer stem cells not only lead to the occurrence of breast cancer, but also may cause breast cancer metastasis and recurrence. The relationship between stem cells and cell resistance is also gaining increasing attentions, and the focus on the stem cell treatment may result in unexpected results. OBJECTIVE: To explore the reversal effect of psoralen on glutathione-S-transferase π (GST-π) in human breast cancer MCF-7/ADR cells and its mechanism. METHODS: MCF-7/ADR cells were cultured and enriched in serum-free medium to obtain breast cancer stem cells. RT-PCR and western blot were used to detect the expression of GST-π at the levels of gene and protein in the MCF-7/ADR cells after treatment with 0, 4, 8, 12, 16 mg/L psoralen. To observe the activation of nuclear factor-κB, western blot was used. The expression of GST-π was detected by RT-PCR in 18 μmol/L SN50 group and 8 mg/L psoralen group. Cell counting kit-8 assay was used to detect the effect of doxorubicin on cell proliferation. RESULTS AND CONCLUSION: Compared with the control group, psoralen reduced the expression of GST-π at the mRNA and protein levels, and significantly inhibited the activation of nuclear factor-κB. It was suggested that psoralen could reverse the multidrug resistance of human breast cancer MCF-7/ADR stem cells by decreasing the expression level of GST-π. The mechanism may be achieved by inhibiting the nuclear factor-κB signal pathway.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of recombinant adenovirus vector expressing human lactoferrin on proliferation and apoptosis of cervical cancer stem-like cells Zhang Xia, Du Zhi-xiang, Wang Na, Meng Ya-li 2017, 21 (13):  2009-2014.  doi: 10.3969/j.issn.2095-4344.2017.13.008 Abstract ( 458 )   PDF (4342KB) ( 220 )   Save BACKGROUND: Gene engineering plays an important role in the process of human malignant tumor treatment, and adenovirus vectors for gene therapy are commonly used. In recent years, lactoferrin is found to exert important effects on the occurrence, development and metastasis of a variety of tumors. However, little is reported on the in vivo expression of adenovirus vector mediated lactoferrin in the human body. OBJECTIVE: To investigate the effect of adenovirus mediated human lactoferrin (Ad-hLF) on proliferation and apoptosis of cervical cancer stem-like cells. METHODS: Primary cervical cancer cells from mice were cultured in vitro to sort cervical cancer stem-like cells using SP method. Afterwards, the stem-like cells were divided into three groups, blank control group, empty vector group, and Ad-hLF group, followed by transfection with nothing, Ad-GFP and Ad-hLF, respectively. After transfection, MTT assay was used to detect the proliferation of cervical cancer stem-like cells; scratch-wound assay was employed to detect the migration of cervical cancer stem-like cells; western blot assay was used to determine the expression of lactoferrin in cervical cancer stem-like cells; and Annexin V-FITC/PI was used to detect cell apoptosis. RESULTS AND CONCLUSION: Compared with the blank control and empty vector groups, Ad-hLF significantly inhibited the proliferation of cervical cancer stem-like cells and reduced the number of scratched cells, while the cell apoptosis increased in the Ad-hLF group (P < 0.05 or P < 0.01). These findings indicate that the recombinant adenovirus vector mediated lactoferrin Ad-hLF for transfection of cervical cancer stem-like cells can be stably expressed in cervical cancer stem-like cells, and can inhibit the proliferation, increase apoptosis and reduce migration of cervical cancer stem-like cells.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biological features and differentiation of human umbilical cord mesenchymal stem cells at different passages Liu Yu-si, Wang Feng-chang, Xia Hai-xiong, Xu Jian-wei, Yang Yan, Cui Dong-bing, Wang Li-long, He Zhi-xu, Shu Li-ping 2017, 21 (13):  2015-2022.  doi: 10.3969/j.issn.2095-4344.2017.13.009 Abstract ( 370 )   PDF (5618KB) ( 384 )   Save BACKGROUND: As a kind of emerging seed cells in the field of tissue engineering and regenerative medicine,  human umbilical cord mesenchymal stem cells are applied depending on whether it can remain stable biological characteristics and multi-directional differentiation capacity after long-term passage in vitro, which needs further experimental studies. OBJECTIVE: To investigate differences of vital biological features of human umbilical cord mesenchymal stem cells over different passages.  METHODS: Human umbilical cord mesenchymal stem cells taken from Wharton's jelly were isolated and cultured using tissue explant method. The proliferation abilities of cells cultured for 3-6 generations were detected using both cell counting method and MTT method. Expressions of surface markers of the cells after expanding over nine passages were comprehensively determined by flow cytometry. Karyotype analysis of cells cultured for 3-6 generations was assessed using cell staining method. The 3rd generation of cells were used to detect differentiation abilities into nerve cells, osteoblasts and hepatocytes.   RESULTS AND CONCLUSION: Proliferation curve of cultured cells was “S” shape, and it was in the slow growth phase within 1-2 days, in the increased logarithmic growth phase within 2-4 days, in the plateau growth phase within 4-5 days, and in the declining growth phase within 5-6 days. Furthermore, two methods of testing results were consistent. Expression levels of cell surface markers CD90, CD44, CD73 were all above 90% while the expression levels of CD11b, CD18, CD34, CD43 and HLA-DR were very low at passages 1-9. No abnormality in cell karyotype was found in cells at passages 3-6. Human umbilical cord mesenchymal stem cells could differentiate into nerve cells, osteoblasts and hepatocytes. All theses experimental findings show that the human umbilical cord mesenchymal stem cells have high proliferation activity and multi-directional differentiation properties.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of cultivation systems on the maintenance of human umbilical cord mesenchymal stem cell characteristics and their proliferation rate in vitro Sun Ya-ru, Zhang Bing-qiang, Wang Fu-bin, Xu Ping, Wang Er-pu, Li Cui-cui 2017, 21 (13):  2023-2028.  doi: 10.3969/j.issn.2095-4344.2017.13.010 Abstract ( 288 )   PDF (6295KB) ( 282 )   Save BACKGROUND: Preliminary data showed that the application of human platelet lysate to human umbilical cord mesenchymal stem cell culture can better maintain the characteristics of stem cells than the application of serum-free medium. However, the serum-free medium can better improve the proliferation of mesenchymal stem cells in vitro than the human platelet lysate. OBJECTIVE: To screen out a better mesenchymal stem cell cultivation system that can greatly maintain the characteristics and proliferation rate of human umbilical cord mesenchymal stem cells. METHODS: Six human umbilical cord specimens were inoculated in six culture systems, and the primary culture of umbilical cord mesenchymal stem cell was performed. These six culture systems were respectively MesenCult serum-free medium with 2% human platelet lysate (group A), StemPro serum-free medium with 2% human platelet lysate (group B), MesenCult serum-free medium (group C), StemPro serum-free medium (group D), low glucose-DMEM with 10% fetal bovine serum (group E), low glucose-DMEM with 10% human platelet lysate (group F). The cells were subcultured at 14 days after inoculation to compare the effects of different culture systems on the morphology, surface markers, differentiation and proliferation of human umbilical cord mesenchymal stem cells. RESULTS AND CONCLUSION: (1) The morphology of passage 3 cells in group D was elongated and uneven in size. The morphology of passage 3 cells was flattened in groups E and F, but the cells in the other groups were spindle-shaped and uniform. There were no significant changes in morphology and size between passage 3 and 5 cells in A and B. In group C, the morphology of passage 5 cells was more flattened and uneven in size compared with passage 3 cells. In group D, the morphology of passage 5 cells was more elongated than that of passage 3 cell. In group E, the morphology of passage 5 cells was more flattened than that of passage 3 cells. There was no significant difference in morphology between passage 3 and 5 cells in group F. (2) The expression rate of cell surface markers had no significant difference at different passages in each group. (3) The adipoinduction and osteoinduction rates were relatively higher in groups A and B compared with groups E and F, and lowest in groups C and D. (4) The cell proliferation rate for each passages in group A was significantly higher than that in group C. The cell proliferation rate for each passage in group B was significantly higher than that in group D. The cell proliferation rate for each passage in groups E and F was significantly lower than that in groups A and B. To conclude, these results suggest that the combination of serum-free medium with human platelet lysate could better maintain the characteristics and the proliferation efficiency of mesenchymal stem cells.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Neural stem cell transplantation effects on neuronal apoptosis, differentiation and neurobehavior changes in rats with cerebral ischemia Liu Yi-min, Zhao Yan-yan, Chen Hong-bing 2017, 21 (13):  2029-2035.  doi: 10.3969/j.issn.2095-4344.2017.13.011 Abstract ( 260 )   PDF (5229KB) ( 236 )   Save BACKGROUND: In recent years, neural stem cell transplantation has become a research hotspot in the treatment of brain injuries, such as cerebral ischemia, but the mechanism has not been fully elucidated. OBJECTIVE: To study the effect of neural stem cell transplantation on neuronal apoptosis and differentiation and neurobehaviors in cerebral ischemia rats. METHODS: Ninety Sprague-Dawley rats with cerebral ischemia were randomized into model group, phosphate buffer solution (PBS) group and neural stem cell group (n=30 per group). Another 30 rats were selected as sham operation group with vascular exposure but with no occlusion under anesthesia. Thirty normal 30 rats acted as controls with no treatment. Thereafter, neurological deficit scores, cerebral infarction volume, the number of apoptotic nerve cells, BrdU/NeuN and BrdU/GFAP expression were detected at 3, 7, 14 days after modeling. RESULTS AND CONCLUSION: On the 7th day and 14th day after modeling, the neurological deficit scores, cerebral infarct volume and the number of apoptotic nerve cells in the neural stem cell group were significantly lower than those in the model group and PBS group (P < 0.05), but there was no statistically significant difference between the model group and PBS group (P > 0.05). On the 3rd day, 7th day and 14th day after modeling, the neurological deficit scores and the number of apoptotic nerve cells in the control group and sham operation group were significantly lower than those in the model group, PBS group and neural stem cell group (P < 0.05). On the 3rd day after modeling, the BrdU/NeuN positive cell count in the model, PBS and neural stem cell groups showed no statistically significant differences (P > 0.05); on the 7th day and 14th day after modeling, the BrdU/NeuN positive cell count in the neural stem cell group was significantly higher than that in the model group and PBS group (P < 0.05), but the difference between the model group and PBS group was not statistically significant (P > 0.05). On the 3rd day, 7th day and 14th day after modeling, BrdU/GFAP positive cell count in the neural stem cell group was significantly lower than that in the model group and PBS group (P < 0.05), and the latter two groups had no statistically significant difference (P > 0.05). In summary, neural stem cell transplantation for cerebral ischemia contributes to the neuronal differentiation of endogenous neural stem cells in the subventricular zone of the lateral ventricles, reduce neuronal apoptosis, and ease ischemic damage to the rat neurological function.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Histological changes of bone marrow mesenchymal stem cells combined with Bio-oss in repairing rabbit skull defects Shao Yan-lin, Luo Shi-jun, Sun Song, Sun Yong, Zhong Ke, Chen Hong-liang 2017, 21 (13):  2036-2042.  doi: 10.3969/j.issn.2095-4344.2017.13.012 Abstract ( 290 )   PDF (2565KB) ( 320 )   Save BACKGROUND: Some studies have focused on bone marrow mesenchymal stem cells (BMSCs) combined with allograft bone or artificial bone substitute materials for bone defect repair. But there is no report on BMSCs combined with Bio-oss for repair of rabbit skull defects as yet. OBJECTIVE: To observe the effect of BMSCs combined with Bio-oss in repairing skull defects in rabbits. METHODS: BMSCs from male rabbits were isolated, cultured, and used as seed cells. In the skull of the female rabbits, three full-thickness bone defects with the same external diameter of 6 mm were made by a ring bone drill. Ninety-six female rabbits were randomly divided into four groups, and given Bio-oss/BMSCs in combination group, Bio-oss alone in Bio-oss group, BMSCs implantation in BMSCs group, and no intervention in blank group. All the implant surfaces were covered with guided tissue regeneration membrane. RESULTS AND CONCLUSION: The osteogenic effect in the combination group was better than that in the other three groups, and the Bio-oss group showed better osteogenesis in comparison with BMSCs and blank groups. But there was no significant difference between the BMSCs and blank groups. These findings indicate that the combined use of BMSCs as seed cells and Bio-oss as a scaffold material exerts overt osteogenic effects in rabbit skull defect area, which provides a new idea for the clinical treatment of bone defects.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of musk ketone on in vivo migration of exogenous bone marrow mesenchymal stem cells in skull defect rats Hou Fei-yi, Xie Xing-wen, Li Shen-song, Shao Hong-bin, Zhang Lian 2017, 21 (13):  2043-2048.  doi: 10.3969/j.issn.2095-4344.2017.13.013 Abstract ( 287 )   PDF (1375KB) ( 265 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are a kind of stem cells with multi-directional differentiation ability and play an important role in the healing of fractures. Therefore, it is of great significance to explore how to promote the BMSCs migration in vivo, thereby promoting bone defect repair. OBJECTIVE: To study the effects of different concentrations of musk ketone on in vivo migration of exogenous BMSCs, and to preliminarily explore the mechanism of fracture healing and cited theory.   METHODS: A rat model of skull defects was made. Passage 3 BMSCs were harvested by using adherence method, labeled with DAPI, and then injected via the tail vein into the model rats. After that, the rats were intragastrically administrated with 0 (blank control), 42 (low dose), 84 (middle dose), 168 μL/kg (high dose) musk ketone, respectively. RESULTS AND CONCLUSION: The number of exogenous BMSCs in the defect region, and the expression of stem cell factor and Fractalkine showed a significant increase in the low- and middle-dose groups compared with the high-dose and blank control groups. These findings indicate that the low- and middle-dose musk ketone can promote the in vivo migration of exogenous stem cells.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Therapeutic effect of autologous bone marrow mesenchymal stem cells on hepatic fibrosis, liver function, MELD score and 1-year survival rate in patients with decompensated hepatitis B Wu Yu-zhuo, Yang Le, Zhai Yu-feng, Zhang Huai-hong, Huo Li-ya 2017, 21 (13):  2049-2055.  doi: 10.3969/j.issn.2095-4344.2017.13.014 Abstract ( 286 )   PDF (1265KB) ( 287 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can differentiate into hepatic stem cells in a specific environment, and participate in the repair and reconstruction of liver function. OBJECTIVE: To investigate the therapeutic effect of autologous bone marrow mesenchymal stem cell transplantation on decompensated hepatitis B. METHODS: Eighty-four patients with decompensated hepatitis B were randomly divided into two groups: normal group (n=42) with symptomatic treatment, and oral entecavir, Fuzheng Huayu Capsule; stem cell group (n=42) with the left and right hepatic artery transplantation of autologous bone marrow mesenchymal stem cells (1×106/kg) based on conventional treatments. Degree of liver fibrosis, liver function and score on Model for End-Stage Liver Disease (MELD) system, 1-year survival rate of patients were detected and analyzed with statistics before and 12, 24 weeks after treatment. RESULTS AND CONCLUSION: (1) Liver fibrosis after treatment in two groups: hyaluronic acid, laminin, type III collagen and type IV collagen levels after treatment were lower than those before treatment in both two groups (P < 0.05), while these indexes in the stem cell group were lower than those in the normal group at 12 and 24 hours after treatment (P < 0.05). (2) Liver function: after treatment, decreased alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels were found in both two groups (P < 0.05), and albumin, cholinesterase, prothrombin activity levels were higher than those before treatment (P < 0.05). The alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels in the stem cell group were lower than those in the normal group at 12 and 24 weeks after treatment, while the cholinesterase level was higher in the stem cell group (P < 0.05). (3) MELD scores: MELD scores were both decreased in the two groups after treatment (P < 0.05), and lower in the stem cell group compared with the normal group at 24 weeks after treatment (P < 0.05). (4) The 1-year survival rate was higher in the stem cell group (69%) than the normal group (50%; P < 0.05). To conclude, the use of autologous bone marrow mesenchymal stem cell transplantation in the treatment of hepatitis B can significantly improve the patients’ liver fibrosis and liver function, and improve the 1-year survival rate of patients.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Therapeutic effect of human umbilical cord mesenchymal stem cells transplantation in the treatment of chronic liver failure Zhang Sui, Liu Li-ping, Ma Wen-bo, Jia Bei, Wang Hai-fang, Xie Cong-jun 2017, 21 (13):  2056-2061.  doi: 10.3969/j.issn.2095-4344.2017.13.015 Abstract ( 287 )   PDF (1180KB) ( 251 )   Save BACKGROUND: Umbilical cord mesenchymal stem cells are plentifully and conveniently obtained with a high proliferative ability, and have opened up a new way to treat patients with liver failure as they can differentiate into hepatocyte-like cells. OBJECTIVE: To observe the safety and efficacy in the treatment of chronic liver failure by transplanting umbilical cord mesenchymal stem cells. METHODS: Using parallel contrast method, 50 patients with chronic liver failure were divided into two groups, namely a stem cell group and a control group, containing 25 patients in each group. For the first group, transplantation of umbilical cord mesenchymal stem cells, (1.4-2.3)×106/kg, 100 mL, was given on the basis of medical comprehensive treatment, while for the second group only simple medical comprehensive treatment was given. The injection was done every 15 days, totally three times. Liver functions, prothrombin activity, Model for End-Stage Liver Disease (MELD) score, clinical symptoms, survival and side effects of the patients were observed before and 2, 4, 12 and 24 weeks after the treatment. RESULTS AND CONCLUSION: Compared with the control group, the albumin level and prothrombin activity were significantly increased in the stem cell group 12 and 24 weeks after treatment (P < 0.05), while the MELD score was significantly decreased in the stem cell group at 4, 12 and 24 weeks after treatment (P < 0.05). There was no significant difference in alanine aminotransferase and total bilirubin levels between the two groups (P > 0.05). Four weeks after treatment, clinical symptoms of the stem cell group improved significantly in comparison with the control group (P < 0.05). During the 24-week follow-up, the survival rate in the stem cell group was significantly higher than that in the control group (P < 0.05). Additionally, there were no adverse reactions and liver cancer associated with the stem cell therapy. Results show that it is safe and effective to treat patients with chronic liver failure through the transplantation of umbilical cord mesenchymal stem cells, and the cell transplantation can significantly improve patients’ survival rate.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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PDX-1 transfected human adipose-derived mesenchymal stem cells: differentiation into islet-like cells and treatment of type 1 diabetes via cell transplantation Shi Guang-jun, Bai Guo-li, Tan Xue-ying, Zhu Ji-jun, Xu Ping 2017, 21 (13):  2062-2067.  doi: 10.3969/j.issn.2095-4344.2017.13.016 Abstract ( 313 )   PDF (5744KB) ( 234 )   Save BACKGROUND: Recent studies have shown that adipose-derived mesenchymal stem cells (ADMSCs) not  only have multilineage differentiation potential, but also exert an important role in blood sugar balance and hormone production. OBJECTIVE: To observe the differentiation potential of human ADMSCs (hADMSCs) into functional islet-like cells and the therapeutic effect of hADMSCs transplantation in diabetic rats. METHODS: PDX-1 gene was transfected into hADMSCs by adenovirus. Cell differentiation and insulin secretion were identified and detected by dithizone staining and ELISA, respectively. Twenty male Sprague-Dawley rats were randomly divided into control group (n=4), diabetes group (n=8) and transplantation group (n=8). Rats in the latter two groups were subjected to making diabetic models by 65 mg/kg streptozotocin injection. Afterwards, rats in the transplantation group were given PDX-1 transfected ADMSCs via the tail vein. RESULTS AND CONCLUSION: At 15 days after transfection, the number of insulin positive cells and insulin secretion were both increased significantly (P < 0.05). Fasting glucose levels in the transplantation group decreased significantly (P < 0.05), while the body weight increased significantly (P < 0.05). In the diabetic group, the fasting glucose level still maintained at a high level, and the body weight of rats was significantly decreased. These results implicated that PDX-1 gene could induce hADMSCs differentiating into functional islet-like cells. PDX-1 transfected ADMSCs transplantation is effective in treating diabetic rats, but the mechanism needs further study.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Reversal effect of transplantation of bone marrow-derived endothelial progenitor cells from the liver fibrosis environment in rats with liver fibrosis Liu Ran, Lan Ling, Liu Bo-wei, Yuan Yuan, Qin Ling-yun, Yu Jing 2017, 21 (13):  2068-2073.  doi: 10.3969/j.issn.2095-4344.2017.13.017 Abstract ( 410 )   PDF (5017KB) ( 226 )   Save BACKGROUND: Theoretically, bone marrow-derived endothelial progenitor cells (EPCs) from liver fibrosis rats could be “filtered” by the pathological environment in vivo. These EPCs would be more adapted to the micro-environment of liver fibrosis, and easier to differentiate into mature endothelial cells participating in the intrahepatic vascular remodeling after transplanted into the liver. OBJECTIVE: To explore the effectiveness of transplantation of bone marrow-derived EPCs from the liver fibrosis environment in liver fibrosis rats. METHODS: Twenty-eight Wistar rats were randomly divided into three groups as follows: normal group (n=8) were injected with olive oil, twice per week; model group (n=10) were infused with carbon tetrachloride at a dose of 3 mL/kg body weight (double doses for the first time), twice per week, and infused with normal saline through the tail vein at 2, 3 and 5 weeks; EPCs transplantation group (n=10) were infused with carbon tetrachloride at a dose of 3 mL/kg body weight (double doses for the first time), twice per week, and infused with EPCs suspension through the tail vein at 2, 3 and 5 weeks. Six weeks after final injection, the angiogenesis, hepatocyte proliferation and pathological changes in the liver tissues were observed. The liver function and coagulation function were tested. RESULTS AND CONCLUSION: (1) The pathological changes of the liver: in the model group, fatty degeneration and hepatocyte necrosis in the liver tissue were serious, inflammatory cells were infiltrated around the portal and central vein, the portal areas expanded, and fibrous tissues overgrew. Compared with the model group, these changes were significantly relieved in the EPCs transplantation group (P < 0.05). (2) The expressions of liver-related proteins: compared with the normal group, the levels of hyaluronic acid, laminin, type III procollagen, vascular endothelial growth factor, epidermal growth factor were significantly increased in the model group (P < 0.05). Compared with the model group, the levels of hyaluronic acid, laminin and type III procollagen were decreased significantly (P  < 0.05), and the levels of vascular endothelial growth factor and epidermal growth factor were increased in the EPCs transplantation group (P  < 0.05). (3) Liver function and coagulation function: compared with the normal group, the liver function and blood blotting function of rats were seriously damaged in the model group (P  < 0.05). Compared with the model group, the liver function and coagulation function were obviously improved in the EPCs transplantation group (P  < 0.05). To conclude, transplantation of bone marrow-derived EPCs from the liver fibrosis environment is effective for liver fibrosis in rats. The mechanism may be associated with the promotion of angiogenesis in the liver.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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The use of placental mesenchymal stem cells to repair the damaged endometrium Niu Ting, Li Ai-bin, Chen Li 2017, 21 (13):  2074-2080.  doi: 10.3969/j.issn.2095-4344.2017.13.018 Abstract ( 386 )   PDF (5370KB) ( 240 )   Save BACKGROUND: Placental mesenchymal stem cells are rich in source and easily obtained, which can differentiate into osteoblasts, nerve cells and liver cells. Additionally, there is no immune rejection and ethical issues in the clinical application. Therefore, placental mesenchymal stem cells are considered to be a good source of adult stem cells. OBJECTIVE: To observe the effect of placental mesenchymal stem cell transplantation in the repair of endometrial lesions in rats. METHODS: Endometrial damage models were established in rats by means of thermal damage, and on the 15th day after modeling, these rat models were randomly divided into four groups (n=10 per group): intrauterine injection of 1 mL of allogeneic placenta mesenchymal stem cell suspension (intrauterine transplantation group), intrauterine injection of the same amount of PBS (intrauterine control group), tail vein injection of 1 mL of allogeneic placenta mesenchymal stem cell suspension (intravenous transplantation group), and tail vein injection of the same amount of PBS (intravenous control group). The female rats experiencing the third estrus after modeling were caged with male rats to observe whether the vaginal plug appeared. The female rats were killed the same day when the vaginal plug was observed, and uterus tissues were taken to detect the number of endometrial glands as well as perform immunohistochemistry and western blot detection. RESULTS AND CONCLUSION: The number of endometrial glands was highest in the intrauterine transplantation group followed by the intravenous transplantation group, and lowest in the two control groups (P < 0.05). The expression of integrin ανβ3 shown by immunohistochemistry and western blot was highest in the intrauterine transplantation group followed by the intravenous transplantation, and lowest in the two control groups (P < 0.05). These findings indicate that the placental mesenchymal stem cell transplantation can repair damaged endometrial tissues in rats to different degrees, by increasing endometrial glands count and improving the endometrial receptivity.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Therapeutic efficacy of unrelated donor peripheral blood versus matched sibling allogeneic hematopoietic stem cell transplantation for high-risk acute leukemia Qiu Da-fa, Xu Xiao-jun, Jian Li, Ren Zhi-juan, Niu Xiao-min, Ye Yong-bin, Guo Xiao-juan 2017, 21 (13):  2081-2086.  doi: 10.3969/j.issn.2095-4344.2017.13.019 Abstract ( 341 )   PDF (1030KB) ( 199 )   Save BACKGROUND: Donor selection for high-risk acute leukemia is still controversial. OBJECTIVE: To compare the therapeutic efficacy of the unrelated donor peripheral blood and matched sibling allogeneic hematopoietic stem cell transplantation for high-risk acute leukemia. METHODS: Total 65 patients with high-risk acute leukemia treated during January 2008 to January 2016 were included, in which 30 patients chose the unrelated donor peripheral blood stem cell transplantation (UD), and other 35 chose the matched sibling allogeneic hematopoietic stem cell transplantation (MS) according to the wishes of patients and their own situation. After treatment, the chi-square test, Kaplan-Meier survival analysis method, and other methods were used to compare the implanted and hematopoietic reconstitution, the occurrence of graft-versus-host disease, relapse mortality and long-term survival between the two groups. RESULTS AND CONCLUSION: The implantation rate, platelet hematopoietic reconstitution time, the incidence of acute and chronic graft-versus-host disease, and its type exhibited no significant differences between the two groups (P > 0.05). The relapse rate, total death rate, and transplant-related mortality rates were 10.0%, 50.0%, and 40.0% in the UD group and 20.0%, 48.6%, and 25.7% in the MS groups, respectively, and the intergroup difference was insignificant (P > 0.05). The expected 2-year cumulative disease-free free survival and overall survival rates were (49.4±9.2)% and (52.6±9.2)% in the UD group and (53.9±8.5)% and (53.9±8.5)% in the MS group, respectively, and the intergroup difference was also insignificant (P  > 0.05). Our experimental findings show that unrelated donor peripheral blood stem cell transplantation can be used as an effective alternative in the absence of sibling donors.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Establishment and identification of pancreatic stem cell strain derived from islets of Kunming mice under feeder layer conditions Cen Yan-hui, Yang Rui, Jia Wei, Li Zhong-hua, Zhong Zhen-guo, Zhong Jing, Bao Juan, He Guo-zhen, Wu Xiao-jun, Li Yi-yi 2017, 21 (13):  2087-2093.  doi: 10.3969/j.issn.2095-4344.2017.13.020 Abstract ( 292 )   PDF (3725KB) ( 227 )   Save BACKGROUND: Until now, little has been reported on establishment of pancreatic stem cell strains and lines, and the purification of pancreatic stem cells is difficult since the cell line establish rate is low. OBJECTIVE: To explore a more rational and effective technique of in vitro separation and continuous passage of pancreatic stem cells, with the hope to establish cell strains and even cell lines and to lay the foundation for the follow-up study of pancreatic stem cells in the treatment of diabetes. METHODS: Firstly, Percoll discontinuous density gradient centrifugation method was applied to separate the mouse pancreatic endocrine portion from the exocrine portion, then to obtain cell strains with highly proliferative ability and low differentiation from pancreatic endocrine portion-the islet. We used mouse embryonic fibroblasts treated with mitomycin C as a feeder layer, for in vitro continuous culture of islet-derived pancreatic stem cells under feeder layer conditions until they were transferred to the 30th passage to establish cell lines. Then pancreatic stem cell line derived from pancreatic islet was detected and identified by a series of tests including growth characteristic test, morphological observation, related molecular marker identification and differentiation characteristic identification. RESULTS AND CONCLUSION: In the continuous process of passage, pancreatic stem cells showed active proliferative ability, and maintained the typical morphological characteristics of stem cells and expression of pancreatic stem cell marker-Nestin. After induction, pancreatic stem cells showed insulin gene expression, reflecting their differentiation potential. Therefore, under the condition of feeder layer, the pancreatic stem cell line derived from Kunming mice was successfully established and the related identification was completed, which lays the foundation for the following research.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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RhoA/ROCK pathway of neural stem cells under mild hypothermia Li Chao, Fu Hong-jie, Zhang Jian-jun, Wang Dong 2017, 21 (13):  2094-2099.  doi: 10.3969/j.issn.2095-4344.2017.13.021 Abstract ( 253 )   PDF (1002KB) ( 218 )   Save BACKGROUND: Mild hypothermia can effectively ease secondary brain and spinal cord injuries, which has a definite protective effect on the central nervous system. Meanwhile, mild hypothermia is conducive to the proliferation, activation and growth of transplanted cells by improving the microenvironment of the injured spinal cord. OBJECTIVE: To investigate whether mild hypothermia intervention can regulate the proliferation and apoptosis of neural stem cells through the RhoA/ROCK pathway. METHODS: The neural stem cell injury model was prepared and randomly divided into two groups: normothermia group and mild hypothermia group. The mild hypothermia group was treated with mild hypothermia [(32.0±0.5) ℃]  for 4 hours. Expression of RhoA, RHOCK, Nogo-A and NgR in neural stem cells was detected by RT-PCR and western blot assay. RhoA/RHOCK positive cells were observed by fluorescence microscope. The intracellular Ca2+ concentration in neurons was measured by laser scanning confocal microscopy. RESULTS AND CONCLUSION: The expression of RhoA, RHOCK, Nogo-A and NgR in neural stem cells at mRNA and protein levels was significantly lower in mild hypothermia group than in the normothermia group (P < 0.05). The intracellular Ca2+ concentration in the neural stem cells was lower in the mild hypothermia group than in the normothermia group (P < 0.05). The number of RhoA/RHOCK positive cells in the brain tissue of rats was significantly lower in the mild hypothermia group than in the in the mild hypothermia group (P < 0.05). To conclude, mild hypothermia regulates the proliferation and apoptosis of neural stem cells by inhibiting the RhoA/ROCK pathway.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Human amniotic epithelial cells: culture technology optimization and biological characteristics Liu Chao, Xu Zhi-guo, Wang Shao-hong, Cheng Hong-ling, Yang Xu-wei, Gong Bo, Liu Yang, Xu Chun-yan 2017, 21 (13):  2100-2107.  doi: 10.3969/j.issn.2095-4344.2017.13.022 Abstract ( 342 )   PDF (1459KB) ( 205 )   Save BACKGROUND: As current studies on isolation, culture and cryopreservation of human amniotic epithelial cells (hAECs) are relatively scattered, it is difficult to form a comprehensive and effective solution to meet the clinical needs of stem cells for transplantation in future. OBJECTIVE: To establish the technology of isolation, culture and cryopreservation of hAECs, and to study the biological characteristics of hAECs. METHODS: Orthogonal method was used to study the effects of different factors on the separation, culture and cryopreservation, and range method was adopted to analyze the data to optimize the separation, culture and cryopreservation. We performed cell primary and passage cultures, morphology observed by microscope, drawn cell growth curve and flow cytometry assay, immunofluorescence staining, hepatocyte like cell differentiation to study the biological characteristics of hAECs. RESULTS AND CONCLUSION: (1) The optimal hAECs separation conditions were as follows: trypsin digestions were conducted at a concentration of 0.25%, four times, once for 20 minutes digestion; optimal conditions of culture were 4×108/L cell seeding density, 10 μg/L epidermal growth factor, 5% serum; optimal conditions of cryopreservation were 1×1010/L cell cryopreservation density, 10% dimethyl sulfoxide, 80% serum. (2) The primary cells were adhered to the wall in 2-3 days, exhibiting irregular polygon, paving stone-like growth. Cell adherence and growth rate were accelerated after subculture, and the growth and proliferation ability of passage 2 cells were not significantly decreased after cryopreservation and resuscitation. (3) Immunofluorescence staining showed that the primary cells strongly expressed SSEA-4 and CK19, but did not express Vimentin, CD45 and HLA-DR. The immunophenotype statistics of the primary and passage 4 cells showed the epithelial mesenchymal transition of hAECs in culture process. (4) Immunofluorescence staining showed that the liver cell marker expression of ALB, CK18 was significantly increased after hAECs were induced to differentiate into hepatocyte-like cells. Glycogen staining revealed glycogen synthesis in hAECs after 3 weeks of induction. To conclude, hAECs are easy to obtain and have strong proliferation ability in vitro, and express surface markers for undifferentiated embryonic stem cells.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Multi-directional differentiation potential of subpopulations of mesenchymal stem cells isolated from human skeletal muscle expressing different myogenic and endothelial markers Zhao Ya-guang, Li Yi, Song Zhuo-yue, Li Guang-heng 2017, 21 (13):  2108-2113.  doi: 10.3969/j.issn.2095-4344.2017.13.023 Abstract ( 449 )   PDF (3594KB) ( 197 )   Save BACKGROUND: Mesenchymal stem cells from human skeletal muscle exhibit multi-directional differentiation potential under the influence of osteogenic proteins such as bone morphogenetic protein 4 (BMP4). But the differentiation of a specific cell subpopulation is not yet clear. OBJECTIVE: To characterize the multi-directional differentiation potential of mesenchymal stem cells from human skeletal muscle based on the expression of different surface markers. METHODS: Four different subpopulations were isolated from the human skeletal muscle by fluorescence-activated cell sorting based on their expression of the myogenic-specific marker CD56 and the endothelial-specific markers CD34 and CD144, including CD56+, CD56+CD34+CD144+, CD34+CD144+, and unsorted groups. Osteogenic differentiation of the four groups of the cells was displayed by Von Kossa staining after the treatment with BMP4 alone or BMP4 plus transforming growth factor β3. Chondrogenic differentiation of these cells was displayed by Alcian blue staining. Bone metabolism was assessed by alkaline phosphatase staining. RESULTS AND CONCLUSION: No significant difference in the bone metabolism was found among four groups after the treatment with BMP4 (P > 0.05). Osteogenic and chondrogenic potentials of the four cell subpopulations were significantly different. Under the same osteogenic induction, the CD56+ cells exhibited strongest potential for osteogenic differentiation; and under the same chondrogenic induction, the CD56+CD34+CD144+ cells exhibited better potential for chondrogenic differentiation than the CD56+ cells. These findings indicate that the osteogenic and chondrogenic potentials are intimately associated with the type of mesenchymal stem cells from human skeletal muscle: the CD56+ cells are closely related to the osteogenic potential, while the CD56+CD34+CD144+ cells have stronger chondrogenic potential.      中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Expression of Sox2 and Oct4 in relation to microvessel density in lung cancer tissues Xiang Bao-li, Qian Hai-hong, Zhang Zhi-lin, Su Jing, Chen Li-ping, Zhang Zhi-hua 2017, 21 (13):  2114-2119.  doi: 10.3969/j.issn.2095-4344.2017.13.024 Abstract ( 314 )   PDF (5041KB) ( 228 )   Save BACKGROUND: High expression of stem cell transcription factors, Sox2 and Oct4, has been confirmed to be closely related to the occurrence of lung cancer. OBJECTIVE: To explore the expression of Sox2 and Oct4 in lung cancer tissues and its correlation with microvascular generation and clinical-pathologic features. METHODS: The expression levels of Sox2 and Oct4 and microvessel density (MVD) in lung cancer tissues (60 cases) and normal tissues (60 cases) were examined by immunohislochemistry. Then, the correlation of the expression of Sox2 and Oct4 and MVD value with clinical-pathologic features in lung cancer was analyzed.  RESULTS AND CONCLUSION: The positive expression of Sox2 and Oct4 was 46.67% (28/60) and 71.67% (43/60) in lung cancer tissues, respectively, while Sox2 and Oct4 were negatively expressed in normal tissues, and there was a significant difference between lung cancer and normal tissues (P < 0.001). The MVD value was 16.22±2.18 in lung cancer tissues, which was significantly higher than that in normal tissues (4.36±2.07) (P < 0.001). Expression of Sox2, Oct4 and MVD value were correlated with TNM stage, differentiation degree, ymph node metastasis, vessel invasion, and liver metastasis (all P < 0.05), but not with the patient’s age and gender (both P > 0.05). The MVD value with Sox2 and Oct4 positive expression was significantly higher than that with Sox2 and Oct4 negative expression (P< 0.001). Spearmen analysis results showed that there was no correlation between Sox2 and Oct4 in lung cancer tissues (r=2.752,P > 0.05). To conclude, the high expression of Sox2 and Oct4 has a high correlation with the initiation and progression of lung cancer, and has positive correlation with the MVD, both of which are significantly concerned with the microvascular generation, invasion and hematogenous metastasis of lung cancer.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Underlying mechanisms and potential of adipose-derived stem cells in the treatment of bronchial asthma Weng Min-hua, Yang Hong-xia, Zhang Jian-yong 2017, 21 (13):  2120-2126.  doi: 10.3969/j.issn.2095-4344.2017.13.025 Abstract ( 378 )   PDF (1009KB) ( 216 )   Save BACKGROUND: Currently, inhaled glucocorticoid is still the classic treatment for asthma. Adult stem cell transplantation has made significant progress in a variety of diseases, and it also provides new insights into the treatment of asthma. OBJECTIVE: To summarize the recent advances in the treatment of asthma with adipose-derived stem cells and related adult stem cells, and to discuss the therapeutic safety of adipose-derived stem cells and possible research directions in asthma therapy. METHODS: Relevant articles published from 2001 to 2016 were searched in PubMed, CBM, CNKI, and Wanfang databases. The keywords were “(adipose-derived stem cells[All Fields]) OR (adipose stem cells[All Fields]) OR (adipose-derived mesenchymal stem cells[All Fields]) AND (asthma[All Fields])” in English and Chinese, respectively. RESULTS AND CONCLUSION: A total of 125 literatures were initially searched, and finally 54 representative papers were selected. Adipose-derived stem cells may reduce airway inflammation, airway hyperresponsiveness, ease collagen deposition and scar tissue formation, promote neovascularization, and reconstruct damaged airways in the mouse asthma model through immune regulation. It is necessary to understand its treatment mechanism of action deeply and comprehensively and carry out genomic analysis before introduction of adipose-derived stem cells as a conventional clinical treatment. In summary, adipose-derived stem cells may be a therapeutic potential for the treatment of airway allergic inflammatory diseases such as asthma.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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A review of human amniotic mesenchymal stem cells in the treatment of acute lung injury Tang Ming-mei, Zhao Jian-jun, Cao Zhi-min, Wen Qiang, He Yue-juan, Chen Ling 2017, 21 (13):  2127-2132.  doi: 10.3969/j.issn.2095-4344.2017.13.026 Abstract ( 252 )   PDF (977KB) ( 173 )   Save BACKGROUND: In recent years, with the in-depth research on the characteristics and functions of stem cells, stem cell research has become a hot research targeting the treatment of a variety of diseases. OBJECTIVE: To summarize the research progress of human amniotic mesenchymal stem cells in the treatment of acute lung injury. METHODS: The PubMed and CNKI databases were searched by computer for relevant literatures published from January 2000 to January 2017. The key words were “mesenchymal stem cells, acute lung injury” in Chinese and English, respectively. RESULTS AND CONCLUSION: As an important member of adult stem cells, human amniotic mesenchymal stem cells not only have the advantages and characteristics of most other stem cells, but also have unique advantages, such as these cells can be extracted from a vast of sources, require only non-invasive operations, and involve no violation of ethics. Human amniotic mesenchymal stem cells can be driven by inflammation reaction and homing to the damaged tissue, and be activated by inflammatory factors to secrete a variety of cytokines in a paracrine fashion, which inhibit neutrophils, macrophages and other immune cell migration and inhibit dendritic cell maturity, and reduce the proliferation of NK cells, B lymphocytes and T lymphocytes, and up-regulate anti-inflammatory cytokines and down-regulate proinflammatory cytokines, and alleviate alveolar inflammatory injury through anti-oxidation and anti-apoptotic processes. Human amniotic mesenchymal stem cells can also assist in the repair or homing to the damaged tissue to directionally differentiate to replace the apoptotic and necrotic cells. All of these characteristics of human amniotic mesenchymal stem cells provide new insight into the treatment of acute lung injury. As acute lung injury is caused by many pathogenic factors and its pathophysiological process is very complicated, human amniotic mesenchymal stem cell transplantation in the treatment of acute lung injury has many issues to be resolved.     中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics