小鼠胚胎干细胞分化为心肌细胞的体外实验  

Differentiation of mouse embryonic stem cells into cardiomyocytes in vitro

作　　者：王芳洁[1] 易岂建[1] 郭鹏飞[1] 

机构地区：[1]重庆医科大学附属儿童医院心脏中心,重庆400014

出　　处：《第四军医大学学报》2009年第9期808-811,共4页Journal of the Fourth Military Medical University

基　　金：高等学校博士学科点专项科研基金(教技发20050631005)

摘　　要：目的:探索提高胚胎干细胞(ESC)体外诱导分化为心肌细胞的实验方法.方法:采用悬滴—悬浮—贴壁三步法(悬滴三步法)和悬浮—贴壁两步法(悬浮两步法),按诱导剂的不同分为Ⅰ组:丹参+5-氮杂胞苷(5-aza)组;Ⅱ组:丹参+维甲酸(RA)组;Ⅲ组:5-aza组;Ⅳ组:丹参组;Ⅴ组:RA组;Ⅵ组:阴性对照组6组进行诱导培养,比较在2种培养方法下各组心肌细胞的分化率.结果:Ⅰ,Ⅲ,Ⅳ,Ⅴ,Ⅵ组采用悬滴三步法培养的心肌细胞分化率分别为(77.78±5.09)%,(66.67±8.82)%,(51.11±1.92)%,(44.44±5.09)%,(23.33±3.34)%;采用悬浮两步法培养的心肌细胞分化率分别为(42.22±6.94)%,(36.67±8.82)%,(25.55±10.71)%,(17.78±5.09)%,(10.00±2.84)%.各组心肌细胞分化率悬滴三步法较悬浮两步法高,差异具有统计学意义(P<0.05).采用悬滴三步法培养Ⅰ组心肌细胞分化率最高可达78%,与其它各组相比较差异具有统计学意义(P<0.05).结论:悬滴三步法联合采用中药丹参与5-aza为诱导剂,可为体外ESC分化为心肌细胞的实验方法提供理论依据.AIM： To investigate a method of inducing embryonic stem cells （ESCs） of mouse high differentiation into cardiomyocytes in vitro. METHODS： Six groups were set up according to inducers added in the culture system of hanging drop culture and suspension culture respectively： group Ⅰ （ salvia mihorrhiza and 5-azacytidine as inducer）, group Ⅱ（salvia miltorrhiza and retinoic acid as inducer）, group Ⅲ（5-azacytidine as inducer）, group Ⅳ （ salvia miltorrhiza as inducer）, group Ⅴ（ retinoic acid as inducer） and group Ⅵ（no inducer）. The rates of differentiation of cardiomyocytes from ESCs were compared between the 6 groups using respectively the two culture systems. RESULTS： The rates of differentiation of group Ⅰ, Ⅲ, Ⅳ, Ⅴ and Ⅵ were （77.78 ± 5.09）%, （66.67±8.82）%, （51.11±1.92）%, （44.44±5.09） % and （23.33 ± 3.34 ）% in hanging drop culture and （42.22±6.94）%, （36.67±8.82）%, （25.55±10.71）%, （ 17.78 ± 5.09） % and （ 10. 00 ± 2.84） % in suspension culture, with statistical difference between the two methods （ P 〈 0. 05 ）.The rate of differentiation of the group Ⅰ was the highest among the 6 groups （P〈0.05）by hanging drop and as high as 78% of the beating embryoid bodies （EBs） were found. CONCLUSION ： The application of traditional Chinese medicinal herb salvia mihorrhiza plus 5-azacytidine as inducers in culture system of hanging drop cultures achieves higher rate of differentiation of cardiomyocytes derived from ESCs in vitro.

关 键 词：胚胎干细胞 分化 心肌细胞 5-氮杂胞苷 丹参 

分 类 号：R363[医药卫生—病理学]