成年兔心耳组织体外培养扩增心肌细胞的生物学特性研究

Biocharacters of cultured cardiomyocytes from the atrial appendage of adult rabbits in vitro

目的 探讨成年兔心耳组织体外培养扩增心肌细胞的生物学特性。方法 选新西兰成年兔 11只 ,分别取少量心耳组织 ,酶解法制成细胞悬液 ,体外培养。于培养 0、12、2 6、4 0d进行细胞计数 ;培养 12、2 6d测定细胞染色体含量 ;培养 0、4 0d ,取部分细胞行免疫组化染色检查 ,计算心肌细胞百分比 ;培养 2 6d ,电镜检查细胞的超微结构 ;培养 12d ,取部分细胞置 - 80℃冷冻保存 ,8周后快速解冻 ,体外继续培养观察。结果 细胞计数分别为 (0 5 4± 0 0 6 )× 10 7、(7 38± 0 73)× 10 7、(5 8 0 0± 16 14 )× 10 7、(2 4 91± 11 86 )× 10 7;DNA含量测定S期细胞分别为 2 2 2 %、5 6 5 % ;免疫组化判定心肌细胞的比例分别为 82 %± 11%、87%± 18% ;电镜下可见心肌细胞肌原纤维的横纹 ;冷冻的心肌细胞快速解冻后 ,体外培养可继续生长扩增。结论 成年兔心耳组织来源的心肌细胞 ,在有限的体外传代培养过程中 ,数目可扩增 ,其亚显微结构和成分基本稳定。

Objective To study the biocharacters of cultured card iomyocytes from atrial appendage of adult rabbits in vitro. Methods Cardiomyocytes from 11 adult New Zealand rabbits were cultured in vitro . The number of cardiomyocytes was counted before and on the 12, 26, 40 day of c ulturing. Examination DNA content of the cells were examined on the 12th and 26th day of the culturing. Percentage of cardiomyocytes was measured by immunocytochemistry before and on the 40th day of culturing. Microconformation of the cells was observed by electron microscope after 26 days of culturing. On the 12th day of culturing, some cells were frozer and preserved at -80℃ and thawed and cultured again 8 weeks later. Results The number of the cultured cells was (0 54±0 06)×10 7, (7 38±0 73)×10 7, (58 00±16 14)×10 7 and (24 91±11 86)×10 7. The DNA content observation showed that S stage cells was acounted for 22 2% and 56 5%; The percentage of cardiomyocytes was 82%±11% and 87%±18%; There was cross striation of the cardiomyocytes under electron microscope; The cells continued to proliferate in culturing after frozen and rapid thawing. Conclusion The cardiomyocytes from atrial appendage of adult rabbits could be proliferated in culturing ex vivo .