中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (11): 2019-2022.doi: 10.3969/j.issn.1673-8225.2012.11.028

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

5-氮胞苷诱导培养P19胚胎癌细胞表达心肌结构蛋白★

王海英   

  1. 河北体育学院运动人体科学系，河北省石家庄市  050041
  • 收稿日期:2011-10-28 修回日期:2012-02-22 出版日期:2012-03-11 发布日期:2012-03-11
  • 作者简介:王海英★，女，1978年生，河北省唐山市人，汉族，2004年河北师范大学毕业，硕士，讲师，主要从事生理生化应用的研究。 misswhy@qq.com

Differentiation of monolayer cultured P19 embryonal carcinoma cells into myocardial cells induced by 5-azacytidine in vitro

Wang Hai-ying   

  1. Department of Human Sports Science, Hebei Institute of Physical Education, Shijiazhuang  050041, Hebei Province, China
  • Received:2011-10-28 Revised:2012-02-22 Online:2012-03-11 Published:2012-03-11
  • About author:Wang Hai-ying★, Master, Lecturer, Department of Human Sports Science, Hebei Institute of Physical Education, Shijiazhuang 050041, Hebei Province, China misswhy@qq.com

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摘要：

背景：少数研究者发现5-氮胞苷在结合P19胚胎癌细胞聚集培养时导致有效的心肌分化，而且在P19胚胎癌细胞单层融合培养时，5-氮胞苷诱导也可致心肌分化。
目的：观察5-氮胞苷诱导下P19胚胎癌细胞是否可以分化成肌细胞及心肌结构蛋白的表达。
方法：培养P19胚胎癌细胞，实验分为2组，诱导组以1 μmol/L 5-氮胞苷诱导4 d，对照组加入同体积的PBS。免疫荧光双标检测各组结蛋白和心肌肌钙蛋白T的共表达；Western blot检测各组细胞的心肌肌钙蛋白T表达。
结果与结论：诱导组检测到结蛋白和心肌肌钙蛋白T在同一细胞相同部位共表达，对照组未见两种蛋白表达和共表达；Western blot检测显示诱导组细胞有心肌肌钙蛋白T表达，对照组无心肌肌钙蛋白T。提示5-氮胞苷可以诱导单层培养P19胚胎癌细胞分化成肌细胞并表达心肌结构蛋白。
关键词：P19胚胎癌细胞；5-氮胞苷；心肌细胞；细胞分化；细胞培养；心肌肌钙蛋白T
doi:10.3969/j.issn.1673-8225.2012.11.028

关键词: P19胚胎癌细胞, 5-氮胞苷, 心肌细胞, 细胞分化, 细胞培养, 心肌肌钙蛋白T

Abstract:

BACKGROUND: Several studies have shown that, during confluent culture and monolayer culture, P19 embryonal carcinoma cells can differentiate into myocardial cells induced by 5-azacytidine.
OBJECTIVE: To observe whether P19 embryonal carcinoma cells can differentiate into myocardial cells induced by 5-azacytidine in vitro, as well as the expression of cardiac structural proteins.
METHODS: P19 embryonal carcinoma cells were divided into two groups: experimental group induced by 1 μmol/L 5-azacytidine for 4 days, and control group induced by phosphate buffered saline. Double immunofluorescence staining technique was used to detect the co-expression of Desmin and cardiac troponin T. Western Blot was used to detect the expression of cardiac troponin T.
RESULTS AND CONCLUSION: Co-expression of Desmin and cardiac troponin T was detected in the same part of the same cells in the experimental group, while no expression and co-expression of the two proteins in the control group. And the cardiac troponin T could be detected in the experimental group, while no expression in the control group. The P19 embryonal carcinoma cells under monolayer culture conditions could differentiate into myocardial cells, and express cardiac structure proteins after 5-azacytidine induction in vitro.

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