Control of cellular proliferation in HeLa‐S3 suspension cultures. Characterization of cultures utilizing acridine orange staining procedures
Control of cellular proliferation in HeLa‐S3 suspension cultures. Characterization of cultures utilizing acridine orange staining procedures
复制标题
利用吖啶橙染色程序控制 HeLa-S3 悬浮培养物中的细胞增殖。
DOI：
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发表时间：
1981
影响因子：
5.6
通讯作者：
L. Dethlefsen
中科院分区：
文献类型：
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作者：
K. Bauer;L. Dethlefsen
在补料和未补料的HeLa‐S3悬浮培养物中详细研究了生长控制。两步吖啶橙子染色和流式细胞术分析表明，两种培养类型的细胞红色荧光（与RNA含量成比例）在指数期和平台期之间下降40-50%。同时评估的细胞绿色荧光（DNA含量）表明，在未补料培养物中，处于平台期的G1-DNA含量的细胞增加，而补料培养物显示过渡期的G1-期细胞短暂增加，随后在平台期恢复至与指数培养物相似的值。在两种培养系统中均观察到3 H-胸苷脉冲标记指数随时间下降。这些数据沿着流式细胞术数据表明，在未补料平台期培养物中存在明显的G1期阻滞，并表明在补料平台期培养物中细胞在细胞周期附近随机阻滞。酸性吖啶橙子染色和流式细胞术分析进一步表明，在平台期未补料培养物中存在静止细胞群，其占总细胞的约34%，由死细胞和活细胞组成。相比之下，补料平台培养物显示约14%的静止，大部分是死细胞。此外，两种培养系统均显示克隆形成指数随时间下降，相对于指数期，平台期的细胞周期通过时间较长。这些发现证实了早期的工作，表明环境对体外哺乳动物细胞的生长控制模式有着深远的影响。
Growth control is investigated in detail in fed and unfed HeLa‐S3 suspension cultures. Two‐step acridine orange staining and flow cytometric analysis indicate declines in cellular red fluorescence (proportional to RNA content) of 40–50% between exponential and plateau phase in both culture types. Cellular green fluorescence (DNA content) assessed simultaneously indicates an increment of cells with G1‐DNA content in plateau phase in the unfed cultures, while fed cultures show a brief increment in G1‐phase cells in the transition phase followed by a recovery in plateau phase to a value similar to that of exponential cultures. Temporal declines in the 3H‐thymidine pulse‐labeling index are observed in both culture systems. These data along with the flow cytometry data indicate a distinct G1‐arrest in the unfed plateau cultures and suggest a random arrest of cells about the cell cycle in fed plateau cultures. Acidic acridine orange staining and flow cytometric analysis furthermore indicate the occurrence of a quiescent population comprising approximately 34% of the total cells and consisting of both dead and viable cells in plateau phase unfed cultures. In contrast, fed plateau cultures show approximately 14% quiescent, mostly dead cells. Also, both culture systems show temporal declines in the clonogenic index and a longer cell‐cycle transit time in plateau phase relative to exponential phase. These findings confirm earlier work which indicates that the environment has a profound influence on the mode of growth control for mammalian cells in vitro.