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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 investigate the biological characteristics of adult rabbit auricle tissue cultured in vitro. Methods Eleven New Zealand adult rabbits were selected, a small amount of atrial appendage were taken, and the cell suspension was made by enzymolysis and cultured in vitro. The cell counts were performed on 0, 12, 2, 4 and 4 days. The cell chromosome contents were measured on day 12 and day 26; the cells were cultured on day 0 and day 04; the sections were harvested for immunohistochemical staining to calculate the percentage of cardiomyocytes; The ultrastructure of the cells was examined by electron microscopy. After cultured for 12 days, some cells were stored at -80 ℃, thawed rapidly after 8 weeks, and cultured in vitro. Results The cell counts were (0 54 ± 0 0 6) × 10 7, (7 38 ± 0 73) × 10 7, (58 0 ± 16 14) × 10 7, (2 4 91 ± 1 86) × 10 7; The content of DNA in S phase was 22.2% and 56.5%, respectively. The percentage of cardiomyocytes determined by immunohistochemistry was 82% ± 11% and 87% ± 18% respectively. Fibril stripes; Frozen cardiomyocytes after rapid thawing, in vitro culture can continue to grow and expand. Conclusion The myocardial cells derived from adult rabbit atrial appendage can proliferate in a limited number of subcultures in vitro, and their sub-microstructure and composition are basically stable.