A simple and economical method in purifying dairy goat luteal cells
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- 作者：Zhonghui Wang1 ; wangzh0917@yahoo.com.cn ; Shulin Chen ; cslin9719@nwsuaf.edu.cn ; csl_1359@163.com ; Hongfei Mo ; Yingxue Huang ; Jinyan Li ; Jianbo Sun ; Lei Liu ; Shantin Zhao
- 关键词：Dairy goat luteal cells ; Percoll ; Purification ; Differential detachment
- 刊名：Tissue and Cell
- 出版年：2013
- 出版时间：August, 2013
- 年：2013
- 卷：45
- 期：4
- 页码：269-274
- 全文大小：1500 K
文摘
As an important cell model, luteal cells are used to study the reproductive cycle and pregnancy maintenance, but there has not yet had a simple and economical method in purifing goat luteal cells. In order to find a good method to isolate and purify the luteal cells from the Guan Zhong dairy goat corpus luteua, we compared the purification efficiency of Percoll density gradient centrifugation method with that of the differential detachment method using trypsin. After using these two methods for isolation, the purified cells were identified by staining for 3¦Â-hydroxy steroid dehydrogenase activity. Cell diameter measurements and cell counting were used to categorize isolated cells from both methods. Cell proliferation activity of purified cells from both methods were studied by Cell Counting Kit-8 for 8 days. The results showed that, after Percoll discontinuous density gradient centrifugation, the purity of luteal cells was 98.2 ¡À 1.2 % in Percoll density layer of 30-40 % . In comparison, the purity of luteal cells isolated in differential detachment by trypsin was 74.3 ¡À 1.8 % . Luteal cells purified from both methods stained positive for 3¦Â-hydroxy steroid dehydrogenase activity, and cells purified by Percoll centrifugation showed a more rapid cell proliferation rate than cells purified by trypsin. In conclusion, this study has demonstrated that Percoll density gradient centrifugation was superior to the method of differential detachment in cell purification efficiency and in maintenance of cell proliferation activity.