Morphological Changes of Porcine Granulosa Cells during in vitro Expansion

IF 3.8 3区农林科学 Q1 VETERINARY SCIENCES Pakistan Veterinary Journal Pub Date : 2020-04-01 DOI:10.29261/pakvetj/2020.010

Hoàng Nghĩa Sơn

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Abstract

Received: Revised: Accepted: Published online: September 15, 2019 October 23, 2019 November 23, 2019 January 22, 2020 This study aimed to assess morphological changes of porcine granulosa cells (pGCs) following in vitro long-term culture. The pGCs from passage 5 showed a dramatic enlargement of cytoplasm (14422.600±1300.704 μm) which was 8.23fold higher than pGCs from primary culture (1752.100±102.244 μm). Nuclear areas were increased from passage 0 (164.990±3.461 μm) to passage 4 (399.514±15.110 μm) and passage 5 (416.326±32.683 μm), respectively. The nucleocytoplasmic ratio of pGCs from passage 0 was 0.089±0.002 which was 2-fold higher than passage 4 (0.045±0.002) and 3-fold higher than passage 5 (0.029±0.002). The diameter of microfilament bundles of pGCs was increased from passage 0 (1.171±0.031 μm) to passage 4 (1.550±0.056 μm) and passage 5 (1.579±0.053 μm), respectively. The weak expression of beta-galactosidase was observed in the several pGCs from passage 2, while pGCs from passage 4 showed the strong activity of beta-galactosidase. The flow cytometry analysis demonstrated that the ratio of apoptotic pGCs was increased through in vitro expansion. These results revealed that pGCs exposed replicative senescent characteristics including the increase of nuclear and cell size, as well as the increase of diameter of actin filament bundles in cytoplasm, and the reduction of nucleocytoplasmic ratio. ©2019 PVJ. All rights reserved

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猪颗粒细胞体外扩增过程中的形态学变化

2019年9月15日2019年10月23日2019年11月23日2020年1月22日本研究旨在评估猪颗粒细胞(pGCs)在体外长期培养后的形态学变化。第5代pGCs细胞质明显增大(14422.600±1300.704 μm)，是原代培养pGCs细胞质增大(1752.100±102.244 μm)的8.23倍。核区由第0通道(164.990±3.461 μm)增加到第4通道(399.514±15.110 μm)和第5通道(416.326±32.683 μm)。第0代pGCs的核质比为0.089±0.002，分别是第4代(0.045±0.002)和第5代(0.029±0.002)的2倍和3倍。pGCs的微丝束直径从0(1.171±0.031 μm)增加到4(1.550±0.056 μm)和5(1.579±0.053 μm)。在传代2的几个pGCs中观察到β -半乳糖苷酶的弱表达，而在传代4的pGCs中则表现出较强的β -半乳糖苷酶活性。流式细胞术分析表明，通过体外扩增，凋亡的pGCs比例增加。结果表明，pGCs表现出细胞核和细胞尺寸增大、细胞质中肌动蛋白丝束直径增大、核质比降低等复制性衰老特征。©2019 PVJ。版权所有

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来源期刊

Pakistan Veterinary Journal 兽医-兽医学

CiteScore

4.20

自引率

13.00%

发文量

审稿时长

4-8 weeks

期刊介绍： The Pakistan Veterinary Journal (Pak Vet J), a quarterly publication, is being published regularly since 1981 by the Faculty of Veterinary Science, University of Agriculture, Faisalabad, Pakistan. It publishes original research manuscripts and review articles on health and diseases of animals including its various aspects like pathology, microbiology, pharmacology, parasitology and its treatment. The “Pak Vet J” (www.pvj.com.pk) is included in Science Citation Index Expended and has got 1.217 impact factor in JCR 2017. Among Veterinary Science Journals of the world (136), “Pak Vet J” has been i) ranked at 75th position and ii) placed Q2 in Quartile in Category. The journal is read, abstracted and indexed internationally.