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人二倍体成纤维样细胞(CF-3)作为研究高尔基体生物发生的模型系统。

Human diploid fibroblast-like cells (CF-3) as a model system for the study of Golgi biogenesis.

作者信息

Dell'Orco R T, Terracio L, Douglas W H

出版信息

J Cell Sci. 1980 Dec;46:279-87. doi: 10.1242/jcs.46.1.279.

Abstract

Human diploid fibroblast-like cells were maintained in an arrested, essentially non-mitotic state for extended periods of time in culture by lowering the serum concentration in the medium from 10 to 0.5%. These arrested cells re-entered the proliferative state when subcultivated in medium containing 10% serum. The morphological distribution and enzyme activities associated with the Golgi complex were examined during growth, arrest, and recovery. Cells grown in medium containing 10% serum possessed a well-developed Golgi complex consisting of parallel arrays of membranes and associated vesicles. Galactosyl transferase activity was highest after 3 days growth (17.5 +/- 5.0 nmol galactose transferred/45 min/mg protein) and declined to 9.8 +/- 3.0 at day 7. When the serum concentration was reduced to 0.5%, Golgi complexes were rarely observed by electron microscopy and galactosyl transferase activity was further reduced into medium containing 10% serum recovered from the arrested state and ultrastructurally resembled cells continuously cultured at the higher serum level. Numerous Golgi complexes reappeared and galactosyl transferase activity increased to 13.0 +/- 3.34 days after subcultivation. These results indicate that the Golgi complex can be experimentally manipulated in human diploid fibroblast-like cells in a manner which may be useful for the study of the biogenesis of this organelle.

摘要

通过将培养基中的血清浓度从10%降至0.5%,人二倍体成纤维细胞样细胞在培养中可长时间维持在静止、基本无有丝分裂的状态。当在含有10%血清的培养基中传代培养时,这些静止细胞重新进入增殖状态。在生长、静止和恢复过程中,对与高尔基体复合体相关的形态分布和酶活性进行了检测。在含有10%血清的培养基中生长的细胞具有发育良好的高尔基体复合体,由平行排列的膜和相关的小泡组成。半乳糖基转移酶活性在生长3天后最高(17.5±5.0 nmol半乳糖转移/45分钟/毫克蛋白质),并在第7天降至9.8±3.0。当血清浓度降至0.5%时,通过电子显微镜很少观察到高尔基体复合体,半乳糖基转移酶活性进一步降低。当细胞从静止状态恢复并在含有10%血清的培养基中培养时,超微结构类似于在较高血清水平下连续培养的细胞。传代培养4天后,大量高尔基体复合体重新出现,半乳糖基转移酶活性增加到13.0±3.3。这些结果表明,高尔基体复合体可以在人二倍体成纤维细胞样细胞中通过实验进行操作,这种方式可能有助于研究该细胞器的生物发生。

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