REGULATION OF HUMAN FIBROBLAST GROWTH-RATE BY BOTH NONCYCLING CELL FRACTION AND TRANSITION-PROBABILITY IS SHOWN BY GROWTH IN 5-BROMODEOXYURIDINE FOLLOWED BY HOECHST-33258 FLOW-CYTOMETRY
REGULATION OF HUMAN FIBROBLAST GROWTH-RATE BY BOTH NONCYCLING CELL FRACTION AND TRANSITION-PROBABILITY IS SHOWN BY GROWTH IN 5-BROMODEOXYURIDINE FOLLOWED BY HOECHST-33258 FLOW-CYTOMETRY
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DOI：
10.1073/pnas.80.10.2951
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发表时间：
1983-01-01
期刊：
影响因子：
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通讯作者：
RABINOVITCH, PS
中科院分区：
文献类型：
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作者：
RABINOVITCH, PS
在5-溴脱氧尿苷存在下生长人二倍体成纤维细胞，然后用Hoechst 33258染料进行DNA特异性荧光的流式细胞术分析，可以定量未循环的细胞比例，以及随后2个细胞周期的G1和G2中的细胞比例。该技术可以快速准确地定量这些细胞的生长分数和G1/S转换速率。人二倍体成纤维细胞在所有群体倍增水平的细胞周期动力学揭示了2个组成部分：周期细胞显示出G1/S转换的概率率，和非周期细胞的可变比例。转换概率（退出G1的速率）和细胞的非循环比例作为血清浓度的函数和作为群体倍增水平的函数而系统地变化。这些数据表明，在给定浓度的有丝分裂原存在下，人二倍体成纤维细胞群体的分裂能力存在潜在的异质性。细胞周期动力学模型必须修改，包括调节生长的变化，在循环细胞的分数，以及从G1退出的速度的变化。
Growth of human diploid fibroblasts in the presence of 5-bromodeoxyuridine, followed by flow cytometric analysis of DNA-specific fluorescence with Hoechst 33258 dye, allows quantitation of the proportion of cells that have not cycled, as well as those in G1 and G2 of 2 subsequent cell cycles. This technique allows rapid and accurate quantitation of the growth fraction and G1/S transition rate of these cells. The cell cycle kinetics of human diploid fibroblasts at all population doubling levels reveal 2 components: cycling cells showing a probabilistic rate of G1/S transition, and a variable proportion of noncycling cells. Both the transition probability (rate of exit from G1) and the noncycling proportion of cells change systematically as a function of serum concentration and as a function of population doubling level. The data suggest the existence of an underlying heterogeneity in the population of human diploid fibroblasts with respect to the capacity to divide in the presence of a given concentration of mitogen. Models of cell cycle kinetics must be modified to include regulation of growth by changes in the fraction of cycling cells, as well as by changes in the rate of exit from G1.