共培养环境中大鼠胚胎中脑细胞诱导人脐带间充质干细胞分化  

Co-culture of hUCMSCs and fetal rat mesencephalic cells induces differentiation of hUCMSCs

作　　者：张蕾[1] 谭雪锋[1] 田美玲[1] 张新化[1] 金国华[1] 

机构地区：[1]南通大学医学院人体解剖学系,江苏省神经再生重点实验室,南通226001

出　　处：《神经解剖学杂志》2013年第4期449-454,共6页Chinese Journal of Neuroanatomy

基　　金：江苏高校优势学科建设工程资助项目(PAPD);南通市科技计划项目(K2010040;BK2012073)

摘　　要：目的:检测人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)在与大鼠胚胎中脑细胞共培养环境中酪氨酸羟化酶(tyrosine hydroxylase,TH)和多巴胺转运蛋白(dopamine transporter,DAT)表达的变化。方法:(1)植块法分离培养hUCMSCs,以细胞免疫化学法鉴定其表面特异性标记物的表达;(2)无菌条件下分离E15大鼠胚胎中脑组织,制成单细胞悬液及中脑组织提取液;(3)用大鼠胚胎中脑组织提取液做培养液,将5-溴脱氧尿嘧啶核苷(bromodeoxyuridine,BrdU)标记的hUCMSCs与大鼠胚胎中脑细胞共培养14 d,细胞免疫化学法检测hUCMSCs中TH和DAT的表达。结果:HUCMSCs高表达CD29、CD44、CD73、CD90和CD105,极低表达CD31和CD45。在与大鼠胚胎中脑细胞共培养14 d后,hUCMSCs中TH阳性表达率为(18.06±2.29)%,DAT阳性表达率为(11.14±2.10)%。结论:用植块法可成功分离并体外培养hUCMSCs,其免疫表型符合间充质干细胞(mesenchymal stem cells,MSCs)的特征。在以大鼠胚胎中脑组织提取液作为培养液,并与大鼠胚胎中脑细胞共培养的诱导条件下,hUCMSCs可部分向多巴胺能神经元分化。Objective： To detect the expression of tyrosine hydroxylase （TH） and dopamine transporter（DAT） in human umbilical cord mesenchymal stem cells （hUCMSCs） in the co-culture environment with fetal rat mesencephalic cells. Methods： （ 1 ） HUCMSCs were isolated and cultured by the explantation technique. The specific surface markers of hUC- MSCs were tested by immunocytochemistry. （2） Under sterile condition, embryonic mesencephalic tissue in El5 rats were isolated and made into single cell suspension and midbrain extract. （3） HUCMSCs labeled with bromodeoxyuridine （BrdU） and fetal rat mesencephalic cells were co-cultured with fetal rat midbrain extract for 14 d, then TH and DAT ex- pressed in hUCMSCs were detected by immunocytochemistry. Results： HUCMSCs high expressed CD29, CD44, CD73, CD90 and CD105, and weakly expressed CD31 and CD45. After being co-cultured with fetal rat mesencephalic cells for 14 d, hUCMSCs could expressed TH（18.06 ±2.29）% and DAT（11. 14±2.10）%. Conclusion： HUCMSCs can be i- solated and cultured in vitro successfully by the explantation technique. The phenotype of these cells is similar to that of mesenchymal stem cells（MSCs）. After being co-cultured with fetal rat mesencephalic cells and treated with fetal rat mid- brain extract, hUCMSCs can partially differentiate into dopaminergic neurons.

关 键 词：人脐带间充质干细胞 胚胎中脑细胞 胚胎中脑组织提取液 共培养 多巴胺能神经元 分化 大鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]