Wittstock Matthias, Rehfeldt Charlotte, Nürnberg Gerd, Renne Ulla, Brück Wolfgang, Mix Eilhard, Zettl Uwe K
Department of Neurology, University of Rostock, Gehlsheimner Strasse 20, D-18147 Rostock, Germany.
J Muscle Res Cell Motil. 2003;24(8):521-6. doi: 10.1023/b:jure.0000009818.91051.30.
Objective of the study was to investigate growth characteristics and susceptibility to apoptosis in different murine muscle cell lines (selected for high body weight, DU-6; randomly mated control, DU-Ks; immortalized myoblast cell line, C2C12). Apoptosis was induced by serum deprivation. At days 4, 5, and 6 of cultivation, protein, DNA and the frequency of apoptotic cells were determined. Until day 4, C2C12 accumulated more DNA and protein compared with DU-Ks and DU-6, while exhibiting a lower percentage of apoptotic cells. Serum deprivation impaired the growth of each cell line. C2C12 continued to accumulate DNA and protein after serum deprivation, whereas reductions, indicative of cell death, were apparent in DU-Ks and DU-6. Serum deprivation did not enhance apoptosis in C2C12. Higher percentages of apoptosis were observed in DU-Ks and DU-6 after 2 days of serum deprivation with greater responsiveness of DU-6 to serum deprivation. The results suggest that cell loss in response to serum deprivation is in part due to induction of apoptosis. C2C12 are less sensitive to sub-optimal culture conditions compared with DU-Ks and DU-6 which are at a closer distance to the in vivo status. Moreover, long-term selection for growth decreases the basic frequency of apoptosis of muscle satellite cells, but increases their susceptibility to apoptosis induction.
本研究的目的是调查不同小鼠肌肉细胞系(选择高体重的DU-6;随机交配对照组DU-Ks;永生化成肌细胞系C2C12)的生长特性和对凋亡的易感性。通过血清剥夺诱导凋亡。在培养的第4、5和6天,测定蛋白质、DNA和凋亡细胞频率。直到第4天,与DU-Ks和DU-6相比,C2C12积累了更多的DNA和蛋白质,同时凋亡细胞百分比更低。血清剥夺损害了每个细胞系的生长。血清剥夺后,C2C12继续积累DNA和蛋白质,而在DU-Ks和DU-6中则出现了表明细胞死亡的减少。血清剥夺并未增强C2C12中的凋亡。血清剥夺2天后,在DU-Ks和DU-6中观察到更高的凋亡百分比,且DU-6对血清剥夺的反应性更强。结果表明,血清剥夺引起的细胞损失部分归因于凋亡的诱导。与更接近体内状态的DU-Ks和DU-6相比,C2C12对次优培养条件不太敏感。此外,长期的生长选择降低了肌肉卫星细胞凋亡的基础频率,但增加了它们对凋亡诱导的易感性。