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首页> 外文期刊>Cell cycle >Morphological changes during replicative senescence in bovine ovarian granulosa cells
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Morphological changes during replicative senescence in bovine ovarian granulosa cells

机译：牛卵巢粒细胞复制衰老期间的形态学变化

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The objective of this study was to evaluate replicative senescence of bovine granulosa cells (bGCs) during in vitro long-term culture. WST-1 assay analysis showed that bGCs proliferation was reduced from primary culture to 14th passage. The several bGCs from the 3rd passage and 7th passage exposed the weak activity of beta-galactosidase, while a strongly positive staining of beta-galactosidase was observed in bGCs from 14th passage. Flow cytometry analysis showed that bGCs were induced to cell cycle arrest at G0/G1 phase through in vitro expansion. TERT transcript expression of bGCs was downregulated from primary culture to 14th passage. The cell and nuclear area of bGCs were dramatically increased from 14th passage to 25th passage. The nucleocytoplasmic ratio of bGCs was dramatically reduced in 22th passage (4.32%) and 25th passage (2.45%), comparing to previous passages: primary culture (10.67%), 7th passage (9.21%), or 14th passage (10.33%). The number of microfilament bundle of bGCs was increased in 22nd passage (67.42 +/- 17.76) and 25th passage (56.31 +/- 22.45). The diameter of microfilament bundle of bGCs in 25th passage was dramatically increased to 1.88 +/- 0.32 mu m comparing to the primary culture (1.15 +/- 0.03 mu m). In this study, we also assessed the nuclear form factor which illustrates the level of nuclear circular form. A reduction of nuclear form factor was observed in bGCs during long-term in vitro expansion. The changes of nuclear form factor were correlated to other senescent characteristics, especially the nucleocytoplasmic ratio.
机译：本研究的目的是在体外长期培养过程中评估牛颗粒细胞（BGCS）的复制衰老。 WST-1测定分析表明，BGCS增殖从原发性培养物降低至第14段。来自第3通道和第7通道的几种BGC暴露了β-半乳糖苷酶的弱活性，而来自第14段的BGC在BGC中观察到β-半乳糖苷酶的强烈阳性染色。流式细胞术分析表明，通过体外膨胀，在G0 / G1相中诱导BGC。将BGCs的TERT转录表达从原发性培养到第14段下调。 BGCS的细胞和核面积从第14段到第25段逐渐增加。 BGCs的核细胞膜比在第22段（4.32％）和第25段（2.45％）中显着降低，与先前的通道相比：初级培养物（10.67％），第7段（9.21％）或第14段（10.33％）。 22nd通道（67.42 +/- 17.76）和第25段（56.31 +/- 22.45）增加了BGC的微纤维束的数量。与初级培养（1.15 +/-0.03μm）相比，第25段中BGCS的微纤维束的直径显着增加至1.88 +/-0.32μm。在这项研究中，我们还评估了说明核圆形形式水平的核形态因素。在长期体外膨胀期间，在BGCS中观察到核形态因子的减少。核形态因子的变化与其他衰老特性相关，尤其是核细胞间质。

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