Kessler-Icekson G, Sperling O, Rotem C, Wasserman L
Exp Cell Res. 1984 Nov;155(1):113-20. doi: 10.1016/0014-4827(84)90772-9.
Primary cultures of newborn rat heart cells were grown for up to 3 weeks in serum-free medium supplemented by insulin, hydrocortisone, transferrin and fetuin. The cells resumed spontaneous beating at 20 h post plating. Mean rates of beating on the second and third day were 79.5 and 94 beats per min, respectively. Cell proliferation occurred during the first 3 days of culture with maximal rates of DNA and protein synthesis on the second day. The highest values of creatine kinase activity were observed on days 2-5 and the three cytoplasmic isozymes, MM, MB and BB, were present in the cultures in proportions similar to those of the newborn heart, indicating stability of the differentiated state of the cells. The relative amount of each isozyme remained unchanged throughout the experiments, MM constituted 70-90% of enzyme activity, MB contributed up to 30% and BB did not exceed 15% of activity. The very low proportion of BB and the lack of increase in this isozyme with age of culture support our earlier morphological observations that non-myocytes do not overgrow the culture.
新生大鼠心脏细胞的原代培养物在添加胰岛素、氢化可的松、转铁蛋白和胎球蛋白的无血清培养基中培养长达3周。细胞在接种后20小时恢复自发搏动。培养第二天和第三天的平均搏动率分别为每分钟79.5次和94次。细胞增殖发生在培养的前3天，第二天DNA和蛋白质合成速率最高。肌酸激酶活性在第2 - 5天观察到最高值，三种细胞质同工酶MM、MB和BB在培养物中的比例与新生心脏相似，表明细胞分化状态的稳定性。在整个实验过程中，每种同工酶的相对含量保持不变，MM占酶活性的70 - 90%，MB贡献高达30%，BB不超过活性的15%。BB的比例非常低且随着培养时间的延长该同工酶没有增加，这支持了我们早期的形态学观察结果，即非心肌细胞不会在培养物中过度生长。
