Bolton W E, Barranco S C
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston 77550.
Am J Hum Genet. 1975 May;27(3):394-409.
The population growth kinetics of human diploid skin fibroblasts derived from cystic fibrosis homozygotes and heterozygotes and from normal subjects were investigated. Our data suggest the following: 1. Population doubling times increase with time in culture, with no significant differences observed among the three genotypes tested, when data were compared at the same subculture times or phases of growth. 2. The fraction of dividing cells in a population decreased with the duration in which the cells were in culture. No significant differences were obtained for cells derived from the three genotypes. 3. Cell cycle times were very similar (18-20 hr) when comparing the normal and cystic fibrosis lines or when comparing cystic fibrosis lines in phases 1 and 2 of growth. 4. No significant variations in population doubling times or growth fractions could be attributed to age or sex of the biopsy donor. 5. Variability in growth fractions and doubling times was minimal through the eighth subculture period but was very great in older cultures (tenth subculture). 6. Changes in growth fractions and doubling times appear to be due to the possibility of "aging" of human diploid fibroblasts in culture rather than to the presence or absence of genes for cystic fibrosis. 7. It is strongly indicated that differences in cell kinetic parameters cannot be used as the basis for differentiation between cells derived from normal or cystic fibrosis genotypes.
对来自囊性纤维化纯合子、杂合子以及正常受试者的人二倍体皮肤成纤维细胞的群体生长动力学进行了研究。我们的数据表明如下:1. 在培养过程中,群体倍增时间随时间增加,当在相同传代时间或生长阶段比较数据时,在所测试的三种基因型之间未观察到显著差异。2. 群体中分裂细胞的比例随细胞培养持续时间而降低。来自三种基因型的细胞未获得显著差异。3. 比较正常和囊性纤维化细胞系时,或比较生长阶段1和2的囊性纤维化细胞系时,细胞周期时间非常相似(18 - 20小时)。4. 群体倍增时间或生长分数的显著变化不能归因于活检供体的年龄或性别。5. 在第八代传代期间,生长分数和倍增时间的变异性最小,但在较老的培养物(第十代传代)中非常大。6. 生长分数和倍增时间的变化似乎是由于培养中的人二倍体成纤维细胞“老化”的可能性,而不是由于囊性纤维化基因的存在与否。7. 强烈表明细胞动力学参数的差异不能用作区分来自正常或囊性纤维化基因型细胞的基础。