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Method to get single cardiomyocyte from embryonic stem cells differentiated in vitro and neonatal rat ventricular tissues
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 10
- DOI:
- 10.3321/j.issn:1000-2790.2008.10.003
- Key Word:
- 小鼠;胚胎;干细胞;心肌/细胞学;细胞培养技术
Abstract： 目的:探讨体外分化的小鼠胚胎干细胞及乳大鼠心室肌组织单个心肌细胞获取方法.方法:用胰酶、胶原酶溶液消化乳大鼠心室肌组织及小鼠胚胎干细胞源心肌细胞,获得细胞悬液经Percoll分离纯化后,再低密度接种富集的心肌细胞,通过光镜观察获得的单个跳动的心肌细胞,采用免疫组织化学和激光共聚焦方法鉴定心肌细胞.结果:乳大鼠心室肌组织及小鼠胚胎干细胞源心肌细胞用胰酶、Dispase酶消化,经Percoll分离纯化后,能获得纯的单个心肌细胞,光镜下单个心肌细胞收缩明显而有力.免疫组织化学和激光共聚焦观察发现单个心肌细胞中有肌原纤维束.结论:使用胰酶、Dispase酶消化后,经Percoll分离纯化可从体外分化小鼠胚胎干细胞及乳大鼠心室肌组织获得活性良好的单个心肌细胞.
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