| 摘要: |
| 为获得高纯度和高存活率的心肌细胞,探索心肌细胞大量培养的条件,并为进一步研究心肌细胞的细胞生物学特性奠定基础,同时为生理学和药理学研究提供细胞来源,试验采用胰蛋白酶顺序消化法及差速贴壁法,从1～2 d龄新生大鼠的心肌组织中分离心肌细胞,观测心肌细胞的形态、结构、搏动状态和反映心肌细胞代谢状况的生化参数。结果表明,分离所得的心肌细胞纯度高,培养第3天就开始自发搏动,后连接成片,进而同步收缩;心肌细胞的亚显微结构包括肌丝、Z线、线粒体和大量糖原颗粒,葡萄糖比消耗率为7.44 nmol/(个·d),乳酸比产率为3.83 nmol/(个·d),乳酸转化率为51%,细胞代谢非常旺盛,说明试验培养条件有利于心肌细胞的气体交换和营养物质的摄取,从而有利于心肌细胞的生长。 |
| 关键词: 心肌细胞 细胞分离 细胞培养 大鼠 |
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| 基金项目:国家863高技术研究与发展计划航天领域项目(2002AA744064)。 |
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| Isolating and culturing of neonatal rat cardiac myocytes |
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| Abstract: |
| This study investigated conditions of isolating and culturing of cardiac myocytes so as to obtain lots of pured cardiac muscle cells and supply a cardiac myocytes model for physiological and pharmacological studies in vitro.1-2-day-old neonatal rat cardiac muscle cells were isolated with enzyme by sequential digestion and pre-plating methods,and the acquired cells were cultured in H-DMEM containing 20% fetal bovine serum.The cell type was identified by H-E and anti-sarcomeric α-actin stains.Cardiac myocytes beating and metabolic indexes,including specific consumption rate of glucose,specific production rate of lactate,lactate transform rate were primarily confirmed.The isolated cardiac myocytes adhered to the bottom of the flasks 24 hours after plating and began to contract spontaneously and by day 3-4 formed synchronously contracting networks.The molar ratios of glucose consumed and lactate produced were 7.44 nmol/(cell·day) and 3.83 nmol/(cell·day) respectively.The molar ratio of glucose consumed to lactate produced was 0.51 mmol/mmol.These demonstrated that metabolic activity of cells was very high and the conditions of culturing were good for cardiac myocytes cultured. |
| Key words: cardiac myocyte cell isolation culture rat |
