TSA对滩羊成纤维细胞作为供体细胞的作用

畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (4): 410-416.doi:

TSA对滩羊成纤维细胞作为供体细胞的作用

李向臣¹，姚雅馨^1，2，梁素丽^1，3，王义鹏^1，4，关伟军¹*，马月辉¹*

1．中国农业科学院北京畜牧兽医研究所，北京 100193; 2.东北林业大学野生动物资源学院，哈尔滨 150040;3.西北农林科技大学动物科技学院，杨凌 712100；4.内蒙古农业大学动物科学与动物医学学院，呼和浩特 010018

收稿日期:2009-05-11 修回日期:1900-01-01 出版日期:2010-04-25 发布日期:2010-04-25
通讯作者: 关伟军

Effect of TSA on Fibroblast Cell as Donor Cell in the Tan Sheep

LI Xiangchen¹, YAO Yaxin ^1,2, LIANG Suli^1,3, WANG Yipeng^1,4,
GUAN Weijun¹*, MA Yuehui¹

1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;2.College of Wildlife Resource, Northeast Forestry University, Harbin 150040, China; 3. College of Animal Science and Technology, Northwest A﹠F University, Yangling 712100, China; 4. College of Animal Science and Animal Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China

Received:2009-05-11 Revised:1900-01-01 Online:2010-04-25 Published:2010-04-25
Contact: GUAN Weijun,MA Yuehui

摘要/Abstract

摘要：

为探索成纤维细胞作为供体细胞的潜能和Trichostatin A（TSA）处理供体细胞的适合浓度，提高克隆胚胎的发育水平。本研究分别利用100、50、25、10 ng·mL-1 TSA处理滩羊成纤维细胞，观察细胞形态，统计细胞活率，利用流式分选技术分析处理前后细胞的周期时相,同时通过间接免疫荧光法检测细胞乙酰化水平，并以经过10~50 ng·mL-1TSA处理的成纤维细胞作为供体细胞，检测其重构胚发育水平。结果发现，当TSA的浓度达100 ng·mL-1时，细胞大量死亡，不同浓度TSA处理细胞24 h后，细胞周期被明显抑制在G0/G1期，乙酰化水平明显高于对照组，其中供体细胞经10 ng·mL-1 TSA处理的克隆胚胎卵裂率和囊胚率明显升高(P<0.05, (85.2±3.4)% vs (68.6±6.7)%, (35.6±5.7)% vs (10.4±8.3)%)。结果表明，经10 ng·mL-1 TSA处理的滩羊成纤维细胞周期同步化明显，乙酰化维持较高水平，重构胚胎发育水平明显提高，适合作为成纤维细胞的处理方法和浓度。

Abstract: In order to investigate the potential of fibroblasts as donor cells, and find out the optimal concentration of TSA used to treat fibroblasts, and further to improve the developmental level of cloned embryos, the fibroblasts were treated with 100, 50, 25, 10 ng·mL-1 TSA, cell morphology and viability were investigated. Flow cytometry were adopted to analyse cell cycle. Acetylation level was detected by indirect immunofluorescence, and the developmental level of cloned embryos was evaluated. Most cells treated with 100 ng·mL-1 TSA were dead, but a significant increase of acetylation level was detected; the cell cycle was arrested at G0/G1 checkpoint with the existence of TSA; generally，there was a higher proportion of cleavages and blastulas (P<0.05, (85.2±3.4)% vs( 68.6±6.7)%, (35.6±5.7)% vs (10.4±8.3)%) when the cells were treated with 10 ng·mL-1 TSA. The results suggested that acetylation level, cell morphology and embryonic developmental level were improved and optimal for the donor cells treated with 10 ng·mL-1 TSA.

李向臣;姚雅馨;梁素丽;王义鹏;关伟军;马月辉. TSA对滩羊成纤维细胞作为供体细胞的作用[J]. 畜牧兽医学报, 2010, 41(4): 410-416.

LI Xiangchen;YAO Yaxin;LIANG Suli;WANG Yipeng;GUAN Weijun;MA Yuehui . Effect of TSA on Fibroblast Cell as Donor Cell in the Tan Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2010, 41(4): 410-416.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/

https://www.xmsyxb.com/CN/Y2010/V41/I4/410