催产素联合乳鼠心肌细胞条件培养液诱导胚胎干细胞分化的作用  

Oxytocin combined with neonatal rat cardiomyocytes-conditioned growth medium enhances differentiation of embryonic stem cells into cardiomyocytes

作　　者：滕继伟[1] 郑强荪[1] 欧东波[1] 郭菁菁[1] 陈瑞[2] 王子宽[1] 

机构地区：[1]第四军医大学唐都医院心脏内科,陕西西安710038 [2]第四军医大学唐都医院整形烧伤科,陕西西安710038

出　　处：《心脏杂志》2010年第4期500-505,513,共7页Chinese Heart Journal

摘　　要：目的:探讨安全高效体外诱导胚胎干细胞(ESCs)向心肌细胞(CMs)分化的途径。方法:ESCs经三步法形成拟胚体(embryoid bodies,EBs)后,按培养基的不同分为4组:乳鼠CMs条件培养液诱导组;催产素(oxytocin,OT)诱导组;混合添加诱导组(乳鼠CMs条件培养液加催产素);对照组(无任何添加)。利用免疫组化染色法检测心肌特异蛋白、RT-PCR检测细胞特殊因子的表达、变时性反应观察细胞对心脏药物的反应及观察细胞的超微结构,对比各组分化后细胞的结构及功能。结果:对照组未观察到细胞跳动。混合添加诱导组、OT诱导组和乳鼠CMs条件培养液诱导组早期分别有(90.30±3.43)%、(21.53±2.69)%和(22.37±6.31)%EBs搏动;中期分别有(92.34±2.65)%、(22.36±2.52)%和(24.15±5.12)%EBs搏动;晚期分别有(83.65±6.27)%、(11.35±2.14)%和(10±4.25)%EBs搏动。免疫组化染色法检测、RT-PCR、变时性反应及电镜观察等的结果提示,混合添加诱导组诱导ESCs分化的效率远高于其余各组。结论:OT、乳鼠CMs条件培养液、OT加乳鼠CMs的条件培养液均具有诱导ESCs分化为CMs的作用,且后者的诱导更具有高效性。AIM： To explore a safe and available approach to induce embryonic stem cells （ESCs） to differentiate into cardiomyocytes （CMs） in vitro. METHODS： Embryoid bodies （EBs） formed from ESCs by three steps were then cultured as four groups： 1） combination group （combined with conditioned medium and OT）, 2） oxytocin （OT） group, 3） conditioned medium group （conditioned medium by neonate rat cardiomyocytes）, and 4） control group （without addition）. Structural and functional properties of these ESC-derived CMs （ESCMs） were then evaluated by immunohistochemical staining, RT-PCR, chronotropic response, and observation under transmission electron microscope. RESULTS： There were no spontaneously beating EBs in the control group. At the early differentiation stage there were, respectively, （90.30±3.43）%, （21.53±2.69）% and （22.37±6.31）% of ESCs-derived EBs in combination group, conditioned medium group and OT group exhibiting spontaneous contractions, whereas the intermediate stage was （92.34±2.65）%, （22.36±2.52）% and （24.15±5.12）%, and the late differentiation stage was （83.65±6.27）%, （11.35±2.14）% and （10±4.25）%. Results from immunohistochemical staining, RT-PCR, chronotropic response and observation under transmission electron microscope all suggested that the differentiation efficiency in the experimental group was much higher than in other groups （P〈0.01）. CONCLUSION： OT is used alone or with conditioned medium by neonate rat cardiomyocytes, or OT combined with conditioned medium for cultivation of neonate rat cardiomyocytes can efficiently induce ESCs to differentiate into functional CMs. However, the differentiation efficiency from OT combined with conditioned medium for cultivation of neonate rat cadiocytes is much more efficient.

关 键 词：胚胎干细胞 催产素 条件培养液 分化 心肌细胞 

分 类 号：R984[医药卫生—药品]