MULTIPLE CELL-CYCLES OCCUR IN RAT HEPATOCYTES CULTURED IN THE PRESENCE OF NICOTINAMIDE AND EPIDERMAL GROWTH-FACTOR
MULTIPLE CELL-CYCLES OCCUR IN RAT HEPATOCYTES CULTURED IN THE PRESENCE OF NICOTINAMIDE AND EPIDERMAL GROWTH-FACTOR
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DOI：
10.1002/hep.1840130105
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发表时间：
1991-01-01
期刊：
影响因子：
13.5
通讯作者：
PITOT, HC
中科院分区：
文献类型：
--
作者：
MITAKA, T;SATTLER, CA;PITOT, HC
在含有10 mmol/L烟酰胺和10 ng/ml表皮生长因子的无血清培养基中，诱导大鼠肝细胞原代培养物的多轮细胞分裂。 每个培养物的细胞在第1天和第5天之间几乎翻了一番。 增殖细胞主要为单核细胞。 培养肝细胞中DNA合成的时间过程表明，H-3-胸苷掺入的峰在接种后60小时和82小时观察到。 细胞的标记指数表明，几乎一半的细胞被标记的H-3-胸苷在48至72小时后的时期。 此外，培养物中约20%的肝细胞在培养48 - 96小时之间启动了第二轮细胞周期，如使用H-3-胸苷和5-溴-2 '-脱氧尿苷连续处理所示。 此外，到培养第4天，约40%和15%的中期分裂分别来自第二轮和第三轮细胞分裂。 第5天培养的肝细胞用白蛋白免疫细胞化学染色，第3天用地塞米松和胰高血糖素诱导酪氨酸转氨酶活性。 此外，电子显微镜照片显示，分裂细胞不仅具有肝线粒体和胆小管样结构的许多特征，而且许多细胞还含有一些大的过氧化物酶体，内部有结晶状的类核。
Multiple rounds of cell division were induced in primary cultures of rat hepatocytes in serum-free medium containing 10 mmol/L nicotinamide and 10 ng epidermal growth factor/ml. Cells per culture almost doubled between day 1 and day 5. The proliferating cells were predominantly mononucleate. The time course of DNA synthesis in cultured hepatocytes showed that peaks of the incorporation of H-3-thymidine were observed at 60 hr and 82 hr after plating. Labeling indices of the cells indicated that almost half the cells were labeled with H-3-thymidine in the periods 48 to 72 hr after plating. In addition, about 20% of the hepatocytes in culture initiated a second round of the cell cycle between 48 and 96 hr in culture, as demonstrated by the use of continuous treatments with H-3-thymidine and 5-bromo-2'-deoxyuridine. Furthermore, by day 4 of culture, about 40% and 15% of metaphases resulted from a second and third round of cell division, respectively. The cultured hepatocytes on day 5 stained with albumin immunocytochemically, and the activity of tyrosine aminotransferase was induced by dexamethasone and glucagon on day 3. In addition, electron micrographs revealed that dividing cells not only had many characteristics of liver mitochondria and bile canaliculus-like structures, but many also contained a few large peroxisomes with internal crystalline nucleoids.