低氧预处理骨髓间充质干细胞耐受缺血缺氧的能力

doi:10.3969/j.issn.2095-4344.2013.49.003

中国组织工程研究 ›› 2013, Vol. 17 ›› Issue (49): 8474-8480.doi: 10.3969/j.issn.2095-4344.2013.49.003

• 骨髓干细胞 bone marrow stem cells • 上一篇下一篇

低氧预处理骨髓间充质干细胞耐受缺血缺氧的能力

徐巧岩¹，夏琳²，王吉昌³

1山东省莱州市人民医院呼吸科，山东省烟台市 261400
2山东大学齐鲁医院整形科，山东省济南市 250012
3山东大学第二医院整形科，山东省济南市 250033

修回日期:2013-09-18 出版日期:2013-12-03 发布日期:2013-12-03
通讯作者: 王吉昌，博士，主治医师。山东大学第二医院整形科，山东省济南市 250033
作者简介:徐巧岩，女，1976年生，山东省莱州市人，汉族，1999年山东医科大学毕业，主治医师。
基金资助:
国家自然科学基金项目青年项目(30900309)*

Hypoxic preconditioning effects on the ability of bone marrow mesenchymal stem cells tolerant to ischemia and hypoxia

Xu Qiao-yan¹, Xia Lin², Wang Ji-chang³

1 Department of Respiratory Medicine, the People’s Hospital of Laizhou, Yantai 261400, Shandong Province, China
2 Department of Plastic Surgery, Qilu Hospital of Shandong University, Jinan 250012, Shandong Province, China
3 Department of Plastic Surgery, the Second Hospital of Shandong University, Jinan 250033, Shandong Province, China

Revised:2013-09-18 Online:2013-12-03 Published:2013-12-03
Contact: Wang Ji-chang, M.D., Attending physician, Department of Plastic Surgery, the Second Hospital of Shandong University, Jinan 250033, Shandong Province, China wangjichangxu@163.com
About author:Xu Qiao-yan, Attending physician, Department of Respiratory Medicine, the People’s Hospital of Laizhou, Yantai 261400, Shandong Province, China qiaoyanx@yahoo.cn
Supported by:
the National Natural Science Foundation of China for the Youth, No. 30900309*

摘要/Abstract

摘要：

背景：有研究表明骨髓间充质干细胞的生理环境氧体积分数低于常规培养用的20%-21%，适度低氧体积分数下，骨髓间充质干细胞表现出促进生长增殖，保持分化潜能的特性。
目的：观察低氧预处理后的大鼠骨髓间充质干细胞的生物学特性，以期为低氧预处理后的骨髓间充质干细胞体内移植治疗寻找体外依据。
方法：将大鼠骨髓间充质干细胞传代接种后置于体积分数1%O₂，5%CO₂的低氧培养箱内培养，以体积分数20%O₂，5%CO₂常氧培养的骨髓间充质干细胞同样封闭作为对照。
结果与结论：MTT检测显示，在低氧条件下培养的经低氧预处理的骨髓间充质干细胞较常氧条件下培养的细胞在贴壁后会经历一个相对较长时间的潜伏期，随后才进入指数生长期，体积分数1%O₂为非致死性的培养条件。锥虫蓝染色显示，封闭无血清条件下，两组细胞存活率均随着时间的推移而降低(P < 0.05)，而常氧组下降更快。流式细胞仪检测显示，低氧组的细胞表面分子标记物CD29(+)，CD90(+)，CD31(-)，CD45(-)的表达与低氧预处理前差异无显著性意义，未向血管内皮细胞分化。实时定量PCR和酶联免疫实验检测结果显示，低氧预处理48 h后，骨髓间充质干细胞中血管内皮生长因子mRNA和蛋白的表达均升高(P < 0.05)。Western-blot检测显示，低氧预处理24-48 h后，骨髓间充质干细胞中缺氧诱导因子1α的蛋白表达明显增多(P < 0.05)。证实，体积分数1% O₂能短暂抑制骨髓间充质干细胞的增殖，但不会产生致死性效应，可以作为低氧预处理的氧体积分数。体积分数1% O₂预处理48 h后，骨髓间充质干细胞的表型未发生明显变化，仍维持其干性。低氧预处理提高骨髓间充质干细胞对缺血缺氧的耐受能力，机制可能与低氧诱导因子1α表达增多有关。

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

全文链接：

关键词: 干细胞, 骨髓干细胞, 骨髓间充质干细胞, 低氧预处理, 低氧诱导因子1α, 血管内皮生长因子, 大鼠, 国家自然科学基金

Abstract:

BACKGROUND: According to the previous studies, it is known that bone marrow mesenchymal stem cells live in the physiological environment of lower oxygen concentration compared with the common culture concentration (20%-21%). Hypoxia can promote bone marrow mesenchymal stem cells proliferation and maintain the characteristics of differentiation potential.
OBJECTIVE: To understand the biological characteristics of bone marrow mesenchymal stem cells after hypoxic preconditioning, and search for in vitro basis for bone marrow mesenchymal stem cells therapy in vivo with hypoxic preconditioning strategy.
METHODS: After passage and inoculation, bone marrow mesenchymal stem cells were cultured in the hypoxic incubator (1% O₂, 5% CO₂). Bone marrow mesenchymal stem cells cultured in the normoxic incubator (20% O₂, 5% CO₂) served as controls.
RESULTS AND CONCLUSION: Compared with bone marrow mesenchymal stem cells cultured under normoxia (20%O₂), bone marrow mesenchymal stem cells cultured under hypoxia (1% O₂) began to display exponential growth phase after a longer incubation period. Hypoxia (1% O₂) was not the lethal condition for bone marrow mesenchymal stem cells. Cell survival rates of both groups reduced over time, while those of bone marrow mesenchymal stem cells cultured under normoxia reduced faster (P < 0.05). Surface markers expression of bone marrow mesenchymal stem cells, including CD29(+), CD90(+), CD31(-), CD45(-), after hypoxic preconditioning did not change greatly when compared with the results before hypoxic preconditioning. The hypoxia preconditioned bone marrow mesenchymal stem cells did not differentiate into vascular endothelial cells. Hypoxia-inducible factor-1α protein expression was promoted after 24 hours to 48 hours hypoxic preconditioning, and there was significant difference between the two time points (P < 0.05). Vascular endothelial growth factor expression increased at both transcriptional and protein levels after 48 hours hypoxic preconditioning (P < 0.05). Culture under hypoxia (1% O₂) does not produce a lethal effect on bone marrow mesenchymal stem cells and can be used for hypoxic preconditioning. Surface markers of bone marrow mesenchymal stem cells after 48 hours hypoxic preconditioning exhibit no great changes and the hypoxia preconditioned bone marrow mesenchymal stem cells maintain the characteristics of stemness. Hypoxic preconditioning can promote the ability of bone marrow mesenchymal stem cells to survive under hypoxia/ ischemic condition. The activation of the hypoxic response signal transduction pathway through hypoxia-inducible factor-1α may explain this.

全文链接：

Key words: anoxia, mesenchymal stem cells, stem cell transplantation, cell hypoxia, oxygen

中图分类号:

R394.2

徐巧岩，夏琳，王吉昌 . 低氧预处理骨髓间充质干细胞耐受缺血缺氧的能力[J]. 中国组织工程研究, 2013, 17(49): 8474-8480.

Xu Qiao-yan, Xia Lin, Wang Ji-chang. Hypoxic preconditioning effects on the ability of bone marrow mesenchymal stem cells tolerant to ischemia and hypoxia[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(49): 8474-8480.

图/表（结果） 6

The growth curve of bone marrow mesenchymal stem cells under hypoxia (1% O₂) and normoxia

Both bone marrow mesenchymal stem cells cultured under hypoxia and normoxia from the fourth passage entered the platform period after a 4-day logarithm multiplication period, while the bone marrow mesenchymal stem cells cultured under hypoxia entered the logarithm multiplication period 1 day later than those cultured under normoxia (Figure 1).

HpBMSCs survival in vitro under hypoxia and serum deprivation condition (Figure 2)

After HpBMSCs and nBMSCs from the fifth passage being sealed into serum-free DMEM (low glucose), the cell deaths of both groups were shown to be time-dependent. The survival rates showed statistically significance between the two groups at the time points of12, 24, 36, 48,72 and 84 hours (P < 0.05). The median survival time also showed statistical significance (P < 0.05).

Surface markers of HpBMSCs

The HpBMSCs from the fifth bone marrow mesenchymal stem cells demonstrated to be positive for CD44 and CD29, negative for CD31 and CD45. The results varied slightly with those of the bone marrow mesenchymal stem cells before hypoxic preconditioning (P > 0.05; Table 1).

Expression of VEGF mRNA and protein

The expression of VEGF mRNA increased after bone marrow mesenchymal stem cells being cultured under hypoxia within 48 hours (Figure 3).The VEGF protein level was found significantly higher in the supernatant from the HpBMSCs group, (214.05±20.24 ) pg/μL, than that from the nBMSCs group, (126.55±16.58) pg/μL (P < 0.05; Figure 4).

Expression of HIF-1α protein

HIF-1α protein expression increased after bone marrow mesenchymal stem cells being cultured under hypoxia within 24 hours (P < 0.05; Figure 5). After 48 hours of hypoxic preconditioning, HIF-1α remained a high level in HpBMSCs. However, there was no significant difference between the two time points

(P > 0.05; Figure 5).

参考文献

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[2] Tse WT, Pendleton JD, Beyer WM, et al. Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation. 2003;75(3):389-397.

[3] Li Y, McIntosh K, Chen J, et al. Allogeneic bone marrow stromal cells promote glial–axonal remodeling without immunologic sensitization after stroke in rats. Exp Neurol. 2006;198(2):313-25.

[4] Hamou C, Callaghan MJ, Thangarajah H, et al. Mesenchymal stem cells can participate in ischemic neovascularization. Plast Reconstr Surg. 2009;123(2 Suppl):45S-55S.

[5] Abdollahi H, Harris LJ, Zhang P, et al.The role of hypoxia in stem cell differentiation and therapeutics. J Surg Res. 2011; 165(1):112-117.

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[7] Toma C, Pittenger MF, Cahill KS, et al. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation. 2002;105(1):93-98.

[8] Jin C, Zhang PJ, Wu XM, et al. Impact of hypoxic preconditioning on apoptosis and its possible mechanism in orthotopic liver autotransplantation in rats. Hepatobiliary Pancreat Dis Int. 2009;8(1):40-45.

[9] Leroux L, Descamps B, Tojais NF, et al. Hypoxia preconditioned mesenchymal stem cells improve vascular and skeletal muscle fiber regeneration after ischemia through a Wnt4-dependent pathway. Mol Ther. 2010; 18(8):1545-1552.

[10] Hu X, Yu SP, Fraser JL, et al. Transplantation of hypoxia-preconditioned mesenchymal stem cells improves infarcted heart function via enhanced survival of implanted cells and angiogenesis. J Thorac Cardiovasc Surg. 2008; 135:799-808.

[11] Yu X, Lu C, Liu H, et al. Hypoxic preconditioning with cobalt of bone marrow mesenchymal stem cells improves cell migration and enhances therapy for treatment of ischemic acute kidney injury. PLoS One. 2013;8(5):e62703.

[12] Wang JC, Xia L, Song XB, et al. Transplantation of hypoxia preconditioned bone marrow mesenchymal stem cells improves survival of ultra-long random skin flap. Chin Med J (Engl). 2011;124(16):2507-2511.

[13] Chang CP, Chio CC, Cheong CU, et al. Hypoxic preconditioning enhances the therapeutic potential of the secretome from cultured human mesenchymal stem cells in experimental traumatic brain injury. Clin Sci (Lond). 2013; 124(3):165-176.

[14] Ohnishi S, Yasuda T, Kitamura S, et al. Effect of hypoxia on gene expression of bone marrow-derived mesenchymal stem cells and mononuclear cells. Stem Cells. 2007;25(5): 1166-1177.

[15] Tsai CC, Yew TL, Yang DC, et al. Benefits of hypoxic culture on bone marrow multipotent stromal cells. Am J Blood Res. 2012;2(3):148-159.

[16] Abdollahi H, Harris LJ, Zhang P, et al. The role of hypoxia in stem cell differentiation and therapeutics. J Surg Res. 2011; 165:112-117.

[17] Yu X, Lu C, Liu H, et al. Hypoxic preconditioning with cobalt of bone marrow mesenchymal stem cells improves cell migration and enhances therapy for treatment of ischemic acute kidney injury. PLoS One. 2013;8(5):e62703.

[18] Hu X, Wei L, Taylor TM, et al. Hypoxic preconditioning enhances bone marrow mesenchymal stem cell migration via Kv2.1 channel and FAK activation. Am J Physiol Cell Physiol. 2011;301(2):C362-372.

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引言

Bone marrow mesenchymal stem cells have many advantages. They have the potential to differentiate along the bone, cartilage, fat and muscle lineage. They cannot only differentiate directly and function as a new cell frame, but also function through the release of many growth factors. Their harvest from autologous donor sites is relatively harmless and eliminates ethical

disputes and other concerns related to graft rejection^[1]. Some studies suggest that human bone marrow mesenchymal stem cells are not antigen-presenting cells and cannot cause activation of the host’s immune system^[2], which means that even allogeneic bone marrow mesenchymal stem cells may be used for transplantation therapies^[3]. They can readily be cultured in vitro to obtain sufficient cells for transplantation. They have been widely used for tissue engineering and regenerative medicine study.

Recently, it is found that mesenchymal stem cells can participate in ischemic neovascularization^[4-5]. One major problem of cell transplantation therapy is that a large number of cells die after being transplanted into the hypoxic sites^[6-7].

Hypoxic preconditioning means that the body or tissue gets tolerance for more serious and even fatal ischemia or hypoxia after one or more short-term, non-lethal hypoxic stimulation^[8]. Hypoxic preconditioning has been found to be an effective strategy for a few types of cells survival under ischemic conditions^[9-13]. Recent studies have found that low oxygen tension is an effective stimulus of many gene expressions^[14-15], which may mimic the physical microenvironment of bone marrow mesenchymal stem cells. Hypoxia can accelerate stem cells proliferation and maintain their multipotency^[16-17]. Hypoxic preconditioning enhances bone marrow mesenchymal stem cell migration^[18-19].

We hypothesized that hypoxic preconditioning bone marrow mesenchymal stem cells with 1% O₂ can lead to their acquiring the ability to survive under hypoxic condition for a long time and to take biological effects. If it is true, hypoxic preconditioning with certain low level oxygen could be a new and feasible strategy for cell therapy of ischemic disease.

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
全文链接：

材料方法

Design

The cytological experiment in vitro.

Time and setting

The experiment was performed from February to August in 2011 at the laboratory of Institute of Dental Medicine, Shandong University, China.

Materials

Animals

Eight male Sprague-Dawley rats were used for the experiment. They were from the Animal Experimental Center of Shandong University of Traditional Chinese Medicine, aged 14 days, weighing (56.5±6.2) g, License No. SCXK(Lu)20110003.

The rats were bred and maintained in a specific pathogen germ-free environment and the experiment was conducted according to the Guidance Suggestions for the Care and Use of Laboratory Animals, formulated by the Ministry of Science and Technology of China^[20].

Reagent and instruments

Methods

Culture of bone marrow mesenchymal stem cells from rats

Bone marrow mesenchymal stem cells were isolated and harvested as previously described^[10]. Eight male Sprague-Dawley rats 2-week-old were used as bone marrow donors. Bone marrow mesenchymal stem cells were cultured in vitro with the Sprague-Dawley rat mesenchymal stem cell growth medium by adherent culture. The medium contained low-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplementing with 10% fetal bovine serum. Bone marrow mesenchymal stem cells were routinely cultured and expanded under normoxia (20% O₂, 5% CO₂).

Surface antigens of the third passage cells were examined by the flow cytometry. The cells were found to be positive for CD44 (99.98%) and CD29 (99.88%) and negative for CD34 (98.94%), and CD45 (98.23%). Osteogenic and adipogenic differentiation could be successfully induced.

The growth curve of bone marrow mesenchymal stem cells under hypoxia (1% O₂)

Bone marrow mesenchymal stem cells from the fourth passage were cultured in 96-hole culture plates at the density of 1×10⁷/L under hypoxia (1% O₂, 5% CO₂) and normoxia (20% O₂, 5% CO₂) in the incubators. The hypoxia was maintained at 1% O₂, 5% CO₂ by infusing a balanced nitrogen gas mixture. At each time point, three paralleled wells in each incubator were selected to be tested by 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay.

Results were collected as the average. Cell growth curves under hypoxia (1% O₂) and normoxia (20% O₂) were generated to assess the influence of hypoxia (1% O₂) on bone marrow mesenchymal stem cells proliferation.

Hypoxic preconditioning of bone marrow mesenchymal stem cells

Bone marrow mesenchymal stem cells from the fifth passage were cultured under hypoxia (1% O₂, 5% CO₂) for 48 hours in the incubator. After this procedure, they were defined as hypoxia preconditioned bone marrow mesenchymal stem cells (HpBMSCs). The bone marrow mesenchymal stem cells from the fifth passage cultured under normoxia (20% O₂, 5% CO₂) were named normoxic bone marrow mesenchymal stem cells (nBMSCs), and they were collected as control samples. The cells and supernatant were collected for real time-PCR, western blot and enzyme linked immunosorbent assay (ELISA) tests.

HpBMSCs survival in vitro under hypoxia and serum-free condition

Part of the HpBMSCs were collected and resuspended by serum-free DMEM (low glucose) at the density of 1×10⁸/L. Then they were transported into the Eppendorf tubes. After being filled with the cellular suspension, 21 tubes of each group were sealed and placed into the incubator at 37 ℃. After 12, 24, 36, 48, 60, 72 and 84 hours respectively, every three tubes in each group were taken out for test randomly.

All the adherent and suspended cells in the three tubes of each group were collected and resuspended. Then the survival rates of the cells were calculated by the Automated Cell Counter with trypan blue exclusion test. nBMSCs were treated with the same method as controls.

Flow cytometry test

HpBMSCs were collected and made into single cell suspension (3×10⁹/L). Cell surface antigens (CD29, CD31, CD45, CD90) were examined by flow cytometry. nBMSCs at the same passage were collected and examined as controls.

Real time PCR detection

The expression of vascular endothelial growth factor (VEGF) mRNA was examined by real time PCR. The total RNA was extracted with RNeasy Mini Kit. The primer sequences were adapted from previous work^[21-22] and were produced by the Boshang Technology (Jinan, China).

The first primer sequence: 5’-GCT TTA CTG CTG TAC CTC CAC-3’; and the second primer sequence: 5’-AGA AGT TCG GCA GGA CAC-3’. After cDNA synthesis, each sample of 25 μL total volume was analyzed in duplicated by Mastercycler^® ep realplex RT-PCR system. Product specificity was verified by melting curve analysis.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA amplified from the same samples served as an internal control. The result of relative expression was analyzed by the method of 2^-^△△^CT.

ELISA detection

To observe the expression of VEGF protein after hypoxic preconditioning, the supernatant was collected at 48 hours. Three independent cultures were randomly selected for the test. The steps were done according to the instructions of the VEGF ELISA kit (R&D, USA). The average result after hypoxic preconditioning was compared with that under normoxia.

Western blot assay

The expression of hypoxia-inducible factor 1α (HIF-1α) in HpBMSCs was determined by western blot assay. The protein lysate was separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis. Gels were then transferred to polyvinylidene fluoride membranes, blocked with Superbloch, and incubated with primary antibodies (Rabbit polyclonal antibody HIF-1α, Abcam) overnight at 4 ℃. Membranes were then washed in Tris-buffered saline, appropriate secondary antibodies (Goat anti-rabbit IgG, Beijing Zhongshan Golden Bridge) added for 2 hours. After washing with Tris-buffered saline S, the substrate and chemiluminescence were added and membranes developed using the digital imaging system (UVP). The grayscales were analyzed by using Image J 1.41 software.

Statistical Analysis

The results are expressed as mean±SD. All data were analyzed using SPSS 11.0 software (SPSS, Inc., USA). Comparisons between two means were performed by Unpaired Student’s t-tests. Multiple comparisons between three groups were performed by analysis of variance. P < 0.05 was considered statistically significant.

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讨论

When compared with the atmosphere, it was found that bone marrow mesenchymal stem cells live in the hypoxic physical microenvironment. The physical oxygen concentrations varies from 2% to 8% according to previous studies^[23]. As shown in our study, the growth curves of bone marrow mesenchymal stem cells under hypoxia and normoxia demonstrated that bone marrow mesenchymal stem cells growth under hypoxia was inhibited for a longer time after incubation, while they both entered the platform period after a 4-day logarithm multiplication period. We could deduce that 1% O₂ is non-lethal condition for BMSCs and can be used for hypoxic preconditioning. In the present study, 1% O₂ (48 hours) culture was selected for hypoxic preconditioning. After being transplanted to the injury sites, bone marrow mesenchymal stem cells live in the hypoxia/ischemic condition for a long time and a lot of cells die^[6-7].During cell therapy, the bone marrow mesenchymal stem cells were usually suspended and transplanted with serum-free culture medium. We applied the sealed serum-free DMEM (low glucose) to imitate the hypoxia/ischemic condition in vivo and study the survival rate of HpBMSCs. The results of the present study demonstrated that hypoxic preconditioning could improve the survival rate of bone marrow mesenchymal stem cells in the sealed serum-free DMEM (low glucose). Hypoxic preconditioning could be used to improve the therapy results by improving the survival rate of transplanted cell. However, hypoxic preconditioning could not change the trend of cell death under hypoxia/ischemic condition for a long time. It means that the transplanted cells need the restoration of oxygen and nutrient supply as soon as possible to survive further longer.

It has been found that bone marrow mesenchymal stem cells can maintain their stemness under a certain low oxygen concentration^[5]. We applied flow cytometry to examine the surface markers of bone marrow mesenchymal stem cells before and after hypoxic preconditioning. Because the hematopoietic stem cells cannot grow adherently, we did not select the anti-CD34 antibody. We preconditioned the bone marrow mesenchymal stem cells under hypoxia, which may introduce the bone marrow mesenchymal stem cells differentiated into vascular endothelial cells.

We selected the anti-CD31 antibody to examine the trend. However, the results showed that the expression of CD29(+), CD90(+), CD31(-) and CD45(-) did not change greatly after hypoxic preconditioning. That means HpBMSCs did not differentiate into vascular endothelial cells, still maintained their stemness and could be used for cell therapy or seed cells of tissue engineering. The mechanism of hypoxic preconditioning on cell protection is mainly studied in the nervous system. There are some factors involved in the mechanism, such as VEGF, SOD, Bcl22 and HSP70. However the primary factor mediating the response is hypoxia-inducible factor-1 (HIF-1), an oxygen-sensitive transcriptional activator. HIF-1 consists of a constitutively expressed subunit HIF-1β and an oxygen-regulated subunit HIF-1α. The HIF-1 subunit is regulated by oxygen sensing and activated under hypoxia. The present study demonstrated that HIF-1α expression increased 24 hours after hypoxic preconditioning and maintained a high level until 48 hours. The expression of VEGF mRNA and protein increased, too. These results back the mechanism that HIF-1α activation promote VEGF expression under hypoxia for cell survivability. Some scholars have transfected the HIF-1α gene into endothelial progenitor cells and found that the proliferation, differentiation and migration of endothelial progenitor cells are increased. The transfected endothelial progenitor cells can promote angiogenesis in vivo when they are transplanted to the ischemic limb. We all know that gene transfection has many shortcomings, which restrict its clinical application. While hypoxic preconditioning is a simple and feasible method to improve bone marrow mesenchymal stem cells survival and promote the VEGF secretion, it may be used for cell therapy for ischemic diseases.

In summary, culture under hypoxia (1% O₂) does not produce a lethal effect on rat bone marrow mesenchymal stem cells and can be used for hypoxic preconditioning. Surface markers of bone marrow mesenchymal stem cells after hypoxic preconditioning under hypoxia (1% O₂) cannot change greatly and maintain the characteristics of stemness. Hypoxic preconditioning can promote the ability of BMSCs to surviveunder hypoxia/ ischemic condition. The activation of the hypoxic response signal transduction pathway through HIF-1α may explain this.

低氧预处理骨髓间充质干细胞耐受缺血缺氧的能力

Hypoxic preconditioning effects on the ability of bone marrow mesenchymal stem cells tolerant to ischemia and hypoxia

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