A kinetic analysis of myogenesis in vitro.
A kinetic analysis of myogenesis in vitro.
复制标题
体外肌发生的动力学分析。
DOI：
10.1083/jcb.52.1.52
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发表时间：
1972-01
影响因子：
7.8
通讯作者：
Stockdale, F E
中科院分区：
文献类型：
--
作者：
O'Neill, M C;Stockdale, F E
描述了鸡肌肉培养物的条件，该条件产生具有广泛的、相对同步的分化的可再现的生长动力学。研究了细胞密度和培养基变化对细胞融合时间的影响。接种后24小时更换培养基的低密度培养物显示出最高的细胞融合率，在10小时内融合细胞从15%增加到80%。这些最佳的培养条件被用来重新研究早期文献中关于肌肉培养的两个问题：（a）通常在一个细胞周期结束时融合的细胞在融合之前是否可以被迫经历另一个细胞周期;（B）在最后一个S期之后多久细胞可以完成融合？在回答第一个问题时，发现如果改变培养基，可以使许多原本会融合的细胞在融合之前经历另一个细胞周期。在第二种情况下，在融合期开始时，用氚化胸苷孵育不同时间的培养物进行放射自显影。这些显示早在孵育期开始后3小时肌管中的标记细胞核。这表明，细胞可以融合早在G1期的开始，并表明，没有一个强制性的退出细胞周期或延长G1期细胞融合和分化之前，在肌发生。
Conditions which yielded reproducible growth kinetics with extensive, relatively synchronous differentiation are described for chick muscle cultures. The effects of cell density and medium changes on the timing of cell fusion were examined. Low-density cultures which received a change of medium at 24 hr after plating show the highest rate of cell fusion, increasing from 15 to 80% fused cells in a 10 hr period. These optimal culture conditions were employed to reexamine two questions from the earlier literature on muscle culture: (a) can cells which normally would fuse at the end of one cell cycle be forced to go through another cell cycle before fusion; and (b) how soon after its final S period can a cell complete fusion? In answer to the first question, it was found that if the medium is changed, many cells which would otherwise fuse can be made to undergo another cell cycle before fusion. In the second case, radioautographs were made from cultures incubated with tritiated thymidine for various times at the beginning of the fusion period. These show labeled nuclei in myotubes as early as 3 hr after the beginning of the incubation period. This indicates that cells can fuse as early as the beginning of the G1 period, and suggests that there is not an obligatory exit from the cell cycle or a prolonged G1 period before cell fusion and differentiation during myogenesis.