[1]庄淼,殷丽,秦时月,等. 组织块半悬浮培养法分离培养大鼠视网膜血管内皮细胞[J].眼科新进展,2015,35(7):605-607.[doi:10.13389/j.cnki.rao.2015.0164]
　ZHUANG Miao,YIN Li,QIN Shi-Yue,et al. Isolation and cultivation of rat retinal vascular endothelial cells by semi suspension tissue culture[J].Recent Advances in Ophthalmology,2015,35(7):605-607.[doi:10.13389/j.cnki.rao.2015.0164]
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 组织块半悬浮培养法分离培养大鼠视网膜血管内皮细胞
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年7期
页码:
605-607
栏目:
实验研究
出版日期:
2015-07-05

文章信息/Info

Title:
 Isolation and cultivation of rat retinal vascular endothelial cells by semi suspension tissue culture
作者:
 庄淼; 殷丽; 秦时月; 邵珺; 谭澄烨; 姚勇
 214023　江苏省无锡市，南京医科大学附属无锡人民医院
Author(s):
 ZHUANG Miao ; YIN Li ;  QIN Shi-Yue ; SHAO Jun ; TAN Cheng-Ye ; YAO Yong
 Wuxi People ’ s Hospital Affiliated to Nanjing Medical University, Wuxi 214023 , Jiangsu Province , China
关键词:
 ; 细胞培养; 大鼠; 视网膜血管内皮细胞; 免疫组织化学; 细胞鉴定
Keywords:
 ; cell culture ;  rat ;  retinal vascular endothelial cell;  immunochemistry;  cell identification
DOI:
10.13389/j.cnki.rao.2015.0164
文献标志码:
A
摘要:
 目的　利用组织块半悬浮培养法分离培养大鼠视网膜血管内皮细胞。方法　分离大鼠视网膜并剪碎，将４～５个碎块半悬浮培养，培养基为含有体积分数１０％胎牛血清并滴加重组牛碱性成纤维细胞生长因子的ＤＭＥＭ／Ｆ１２培养液，获得的细胞用Ⅷ因子相关抗体鉴定，并使用传统冻存及复苏的方法保存。结果　２４ｈ组织块生长固定在培养皿中，６ｄ可见细胞从组织块边缘游出，并可见多个细胞集落，１６ｄ基本铺满培养皿。Ⅷ因子相关抗体染色阳性，细胞可持续生长并传代。结论　用组织块半悬浮培养法可以简便的获得视网膜血管内皮细胞，所获细胞性状稳定，可用于进一步实验研究。
Abstract:
 Objective To improve the traditional isolation and cultivation methods of rat retinal vascular endothelial cell ( RVEC) by semi suspension tissue culture. Methods The retina isolated from a SPF SD rat was cut into pieces ,4 - 5 pieces were cultured with semi suspension tissue culture method. The 100 mL . L -l fetal bovine serum and recombinant bovine basic fibroblast growth factor were composed into the endothelial cell culture medium. Cultured vascular endothelial cells were identified by factor Vlfl related antigen polyclonal antibody , and preserved by the traditional cryopreservation and resuscitation method. Results The pieces of tissue fragments attached to the petri dish after 24 hours of semi suspension. Then some RVEC travelled from them at 6 days , and the cells in primary culture began to clone . and a cell monolayer was seen at 16 days after culture. RVEC were positive response to FactorⅦ. RVEC could be maintained and passaged. Conclusion With the improved method, the rat RVEC can be isolated more conveniently and effectively. The cells with perfect activity can be used for further research.

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备注/Memo

备注/Memo:
 国家自然科学基金资助（编号：８１４００４１５）；江苏省自然科学基金资助（编号：ＳＢＫ２０１２２２０７３）
更新日期/Last Update: 2015-07-03