心肌细胞培养纯化与小鼠骨髓间充质干细胞分化诱导的研究

发布时间：2018-01-05 11:37

本文关键词：心肌细胞培养纯化与小鼠骨髓间充质干细胞分化诱导的研究　出处：《佳木斯大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的:探讨心肌细胞分离、纯化和培养方法;研究骨髓间充质干细胞(BMSCs)在不同心肌细胞微环境作用下,向心肌样细胞分化能力。方法:采用不同时间点,通过差速贴壁纯化、培养新生乳鼠心肌细胞。将原代乳鼠心肌细胞与BMSCs联合培养,分为:接触性联合培养组,将心肌细胞与BMSCs按2㑳1的比例种植在同一培养皿中;非接触性联合培养组,将分别种植有心肌细胞、BMSCs的盖玻片按2㑳1的比例种放入同一培养皿中共同培养;对照组,BMSCs常规培养。二周后检测各实验组细胞中,心肌特异性蛋白肌钙蛋白T(cTnT)表达情况。结果:心肌细胞培养,采用差速贴壁60min组与90min组之间有统计学意义,差速贴壁60min组、90min组与差速贴壁120min组两两之间有统计学差别。免疫细胞化学鉴定:(1)接触性联合培养组细胞均表达肌钙蛋白T(cTnT);(2)非接触性联合培养组细胞均表达肌钙蛋白T(cTnT);(3)对照组细胞不表达肌钙蛋白T(cTnT)。结论:心肌细胞培养过程中,采用差速贴壁60min较差速贴壁90min、120min获得的心肌细胞数量高;接触性联合培养组与非接触性联合培养组均可将BMSCs诱导分化为心肌样细胞,对照组BMSCs无分化。
[Abstract]:Aim: to investigate the methods of isolation, purification and culture of cardiomyocytes, and to study the ability of bone marrow mesenchymal stem cells (BMSCs) to differentiate into cardiomyocyte-like cells under different microenvironment. Methods: neonatal neonatal rat cardiomyocytes were cultured at different time points by differential adherent purification. Primary neonatal rat cardiomyocytes were co-cultured with BMSCs and divided into contact co-culture group. The relationship between cardiomyocytes and BMSCs was 2? 1% was planted in the same culture dish, and in the non-contact co-culture group, the cover glass of BMSCs implanted with cardiomyocytes was 2? (1) the proportion of BMSCs was cultured in the same culture dish, and the control group BMSCs were cultured routinely. Two weeks later, the expression of cardiac troponin TnT was detected in the cells of each experimental group. Results: there was significant difference between 60 min and 90 min groups in the culture of cardiomyocytes. There was significant difference between 90 min group and 120 min differential adhesion group. (2) the cells in the non-contact co-culture group all expressed troponin TnTnT; 3) the control group did not express troponin TnTnT. Conclusion: in the process of cardiomyocyte culture, the number of cardiomyocytes obtained by differential adhesion for 60 min or 90 min or 120 min is high. Contact co-culture group and non-contact co-culture group could induce BMSCs to differentiate into cardiomyocyte-like cells, while BMSCs in control group had no differentiation.
【学位授予单位】：佳木斯大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329

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