Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/44946
標題: Cell cycle stage analysis of rabbit foetal fibroblasts and cumulus cells
作者: Liu, C.T.
朱志成
Yu, K.C.
Ju, J.C.
關鍵字: nuclear transfer
mammalian-cells
granulosa-cells
in-vitro
embryos
oocytes
differentiation
cloning
serum
mice
期刊/報告no:: Reproduction in Domestic Animals, Volume 39, Issue 6, Page(s) 385-390.
摘要: Synchronization of the cell cycle stages in G(0)/G(1) phase is one of the key factors determining the success of nuclear transplantation. Serum deprivation, contact inhibition and chemical inhibitors are widely used methods for this purpose. In this study, cell cycle stages of foetal fibroblasts and cumulus cells were determined using flow cytometry [fluorescence-activated cell scan (FACS)]. Foetal fibroblasts (in vitro cultured for 72-120 h) and fresh cumulus cells were analysed in Experiment 1. Fifty to 55% proliferating fibroblasts remained in G(0)/G(1) phase compared with 78% in confluent culture (p < 0.05). In contrast to foetal fibroblasts, fresh cumulus cells maintained 90% of the population in the G(0)/G(1) stage. When serum was retrieved from the proliferating fibroblasts from day I to day 5 (Experiment 2), proportions of G(0)/G(1) cells increased from the initial ratio of 53 to 87% at day 4 of starvation, which was significantly higher than the non-starved proliferating cells (p < 0.05). In Experiment 3, fibroblasts were treated with aphidicolin (0.1 mug/ml, 6 h), demicolcine (0.5 mug/ml, 10 h). or a combination of these two chemicals to synchronize the cell cycle stages. Surprisingly, no differences or significantly lower in the proportions of G(0)/G(1) phase cells were detected (25-50%) compared with the uncontrolled growing cells (53%). These results suggested that fresh cumulus cells rest their cell cycle in G(0)/G(1) stage. Serum deprivation became effective in the first 24 h and reached the highest proportions during days 4-5 after deprivation. Chemical synchronization of the cell cycle stage of rabbit foetal fibroblasts to G(0)/G(1) phase appeared less effective compared to serum deprivation.
URI: http://hdl.handle.net/11455/44946
ISSN: 0936-6768
文章連結: http://dx.doi.org/10.1111/j.1439-0531.2004.00525.x
Appears in Collections:動物科學系

文件中的檔案:

取得全文請前往華藝線上圖書館



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.