低氧预处理人羊膜间充质干细胞的生物学特征  

Biological characteristics of hypoxic preconditioned human amniotic mesenchymal stem cells

作　　者：代敏 王帅 张霓霓 黄桂林 余丽梅 胡小华 易杰 姚礼 张立刚 Dai Min;Wang Shuai;Zhang Nini;Huang Guilin;Yu Limei;Hu Xiaohua;Yi Jie;Yao Li;Zhang Ligang(Department of Oral and Maxillofacial Surgery,the Affiliated Stomatological Hospital of Zunyi Medical University,Zunyi 563003,Guizhou Province,China;Key Laboratory of Cell Engineering in Guizhou Province,the Affiliated Hospital of Zunyi Medical University,Zunyi 563003,Guizhou Province,China)

机构地区：[1]遵义医科大学附属口腔医院口腔颌面外科,贵州省遵义市563003 [2]遵义医科大学附属医院贵州省细胞工程重点实验室,贵州省遵义市563003

出　　处：《中国组织工程研究》2021年第19期3004-3008,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81760201),项目负责人:黄桂林;国家自然科学基金(81860198),项目负责人:张霓霓;遵义医科大学硕士启动基金(KY2019-1),项目负责人:代敏。

摘　　要：背景:干细胞移植治疗为许多疑难疾病带来了希望,但移植细胞存活数量少及分泌功能差,很难发挥治疗效用,致使该技术在临床应用受阻。低氧预处理可有效改善植入细胞的增殖活性、抗凋亡能力、分泌功能等生物学特性,因而,低氧预处理的人羊膜间充质干细胞有望在组织损伤修复中发挥更好的治疗作用。目的:探讨低氧预处理对人羊膜间充质干细胞生物学特征的影响。方法:利用机械法和酶消化法分离人羊膜间充质干细胞并传至第3代,将其低氧预处理(体积分数为3%O2)48 h,设置常氧(体积分数为21%O2)对照组,通过CCK-8及Edu增殖实验检测两组人羊膜间充质干细胞增殖活性及DNA合成率。将两组细胞用无血清培养基培养24 h后,Annexin V-FITC/PI实验检测细胞凋亡情况,收集上清液,采用血管生成蛋白芯片(Proteome Profiler)检测上清液中分泌的血管生成相关因子水平。结果与结论:低氧预处理不影响人羊膜间充质干细胞的干细胞特性,能提高人羊膜间充质干细胞的增殖活性;在无血清培养条件下,低氧预处理使人羊膜间充质干细胞的抗凋亡作用得到增强,同时分泌了更多量的血管相关生成因子。BACKGROUND:Stem cell transplantation has brought hope for many difficult diseases,but the survival number of transplanted cells is small and the secretion function is poor,so it is difficult to play a therapeutic effect,which hinders the clinical application of stem cell transplantation.Hypoxic preconditioning can effectively improve the biological characteristics of implanted cells,such as proliferation activity,anti-apoptotic ability and secretion function.Therefore,human amniotic mesenchymal stem cells pretreated with hypoxia may play a better therapeutic role in tissue damage repair.OBJECTIVE: To study the effects of hypoxic preconditioning on the biological characteristics of human amniotic mesenchymal stem cells.METHODS: The mechanical method and enzymatic digestion were used to separate human amniotic mesenchymal stem cells, and then cultured it up to thethird generation. After having hypoxic pretreatment (3% O2) for 48 hours, the nomoxic (21% O2) control group was set. The proliferation activity and DNAsynthesis rate of human amniotic mesenchymal stem cells in the two groups were detected by CCK-8 and Edu proliferation experiments. The two groups ofcells were cultured by serum-free medium for 24 hours. Apoptosis was detected using Annexin V-FITC/PI assay. The supernatant was collected. The levels ofangiogenic factors secreted in the supernatant were detected by using the Proteome Profiler.RESULTS AND CONCLUSION: Hypoxic preconditioning does not affect the characteristics of human amniotic mesenchymal stem cells, but can improve theproliferation activity of human amniotic mesenchymal stem cells;in serum-free culture conditions, hypoxic preconditioning can enhance the anti-apoptoticeffect of human amniotic mesenchymal stem cells, and secrete more angiogenic factors.

关 键 词：干细胞 人羊膜间充质干细胞 低氧 预处理 生物学特征 增殖 凋亡 血管生成 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]