优化细胞因子诱导胎盘源间充质干细胞向胰岛β样细胞分化的实验研究  

Optimization of inducing placenta-derived mesenchymal stem cells to differentiate into islet β-like cells by cytokines

作　　者：陆琰 王娟[2] 吴韶清 张洹[2] LU Yan WANG Juan WU Shao-qing ZHANG Yuan(Guangzhou Cord Blood Bank, Guangzhou Women and Children＇s Medical Center, Guangzhou, Guangdong 510623 Institute of Hematology, Medical College of Jinan University, Guangzhou, Guangdong 510632, China)

机构地区：[1]广州市妇女儿童医疗中心广州脐血库,广东广州510623 [2]暨南大学医学院血液病研究所,广东广州510632

出　　处：《热带医学杂志》2017年第8期1000-1003,1011,F0004,共6页Journal of Tropical Medicine

基　　金：国家自然科学基金(81102248);广东省科技计划项目(2014A020212027)

摘　　要：目的优化多种细胞因子联合诱导人胎盘源间充质干细胞分化为胰岛β样细胞的实验条件。方法从胎盘中分离、扩增间充质干细胞;扩增后的细胞采用激活素A、表皮生长因子(EGF)等多种细胞因子联合诱导;在诱导过程中运用消化重悬、调节葡萄糖浓度等方法优化诱导方案,促进细胞形成胰岛样细胞团,向胰岛β样细胞的分化;比较不同诱导条件下,诱导后的细胞在胰岛素、C肽分泌及胰岛素释放实验中的差异,筛选出最佳方案。结果经诱导后的细胞,免疫荧光染色及免疫细胞化学染色均能检测出PDX-1及胰岛素表达;优化方案3诱导后的细胞不仅胰岛素分泌量明显提高至(222.00±38.00)mU/L,还能分泌C肽至(0.45±0.22)ng/mL,而且对胰岛素释放实验敏感,胰岛素刺激释放指数为(5.93±0.79)。结论经多种细胞因子联合诱导后,人胎盘源间充质干细胞可分化为胰岛β样细胞,胰岛样细胞团的形成有助于胰岛β样细胞的分化发育。Objective To optimize the experiment conditons of inducing human placenta-derived mesenchymal stem cells to differentiate into islet β-like ceils by mutiple cytokines. Methods Mesenchymal stem cells were isolated from placenta and expanded in vitro. Then they were induced by activin A, epidermal growth factor （EGF） and other cytokines. During the induction, cells were digested and resuspended and glucose levels of media were adjusted, which could promote cells to form islet-like clusters and differentiate into islet β-like cells. The insulin content of culture supernatant, C peptide content of culture supernatant and the difference of insulin challenge test were compared among different experimental conditions. And the best experiment conditions were selected, Results After induction, PDX-1 and insulin could be detected by immunofluorescence staining or immunocytochemistry. The insulin secretion level of ceils increased significantly to （222.00± 38.00） mUlL which were induced by the optimized group 3. Moreover, these ceils could secret C peptide （0.45±0.22） ng/mL and were sensitive to insulin release test.The insulin stimulation index of group 3 was （5.93±0.79）. Conclusion These results showed human placenta-derived mesenchymal stem cells could be induced to differentiate into islet β-like cells by multiple cytokines. Formation of islet-like clusters could contribute to the differentiation and development of islet β-like cells.

关 键 词：胎盘 间充质干细胞 分化 胰岛 糖尿病 

分 类 号：Q813[生物学—生物工程]