中国组织工程研究与临床康复2011，Vol.15Issue(1)：92-95,4.DOI:10.3969/j.issn.1673-8225.2011.01.020
悬滴三步法体外诱导小鼠胚胎干细胞向心肌样细胞的分化
Differentiation of mouse embryonic stem cells into cardiomyocytes in hanging drop culture system in vitro
刘官信 1王芳洁 1赵丹 1芦起1
作者信息
- 1. 重庆医科大学儿童医院儿科研究所,儿童发育疾病研究省部共建教育部重点实验室,重庆市,400014
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摘要
Abstract
BACKGROUND: It is theoretical and clinical significance of studying the condition and technique of the differentiation of embryonic stem cell sembryonic stem cell(ESC )into cardiomyocytes and to further elevate rate of differentiation and cell purity.OBJECTIVE: To set up a model of inducing ESC into cardiomyocytes and to provide a suitable method to study development of cadiomyocytes in vitro.METHODS: Undifferentiated ESC were incubated in vitro by hanging drop-suspension-adherence method (hanging drop culture),then divided into six groups according to different inducers added in the culture system: group Ⅰ (salvia miltorrhiza linked with 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). After inducer addition, Cardiomyocyte marker proteins expression of α-actinin, myosin, and α-actin were detected by immunocytochemistry in the ninth day.RESULTS AND CONCLUSION : Spontaneously contracting cells appeared in thesecond day following inducer addition under inverted microscope and one embryoid body had one or more contraction foci. The immunocytochemistry showed the cardiomyocyte marker proteins expression of α-actinin, myosin, α-actin were positive. The rate of differentiation of group Ⅰwas highest, which showed significant difference compared with groups Ⅱ and Ⅵ (P ＜ 0.01), and exhibited significant difference compared with groups Ⅲ, Ⅳ and Ⅴ (P ＜ 0.05). it suggested that traditional Chinese drug salvia miltorrhiza linked with 5-azacytidine as inducers in culture system of hanging drop cultures can improve rates of ESC into cardiomyocytes differentiation in vitro.关键词
胚胎干细胞/分化/心肌细胞/5-氮杂胞苷/丹参/小鼠分类
医药卫生引用本文复制引用
刘官信,王芳洁,赵丹,芦起..悬滴三步法体外诱导小鼠胚胎干细胞向心肌样细胞的分化[J].中国组织工程研究与临床康复,2011,15(1):92-95,4.
