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人结膜上皮细胞的培养鉴定及液氮冻存
引用本文:郑健樑,卢蓉,张洁,林明楷,林建贤.人结膜上皮细胞的培养鉴定及液氮冻存[J].眼科学报,2000,16(2):135-138.
作者姓名:郑健樑  卢蓉  张洁  林明楷  林建贤
作者单位:中山医科大学中山眼科中心,广州 510060
摘    要:目的:探索人眼结膜上皮细胞体外培养和细胞保存的最佳方法,建立人结膜上皮细胞,进一步研究人结膜的病理、生理特点及为毒理试验提供可靠的细胞模型。方法:分别用组织块培养法、机械分离法及混合消化液培养法体外培养正常成年人结膜上皮细胞,通过观察细胞形态、生长特性并用原位免疫组化方法鉴定培养细胞;收集第3代和第4代融合的细胞液氮冻存,保存30天后复苏,观察复苏成功率。结果:3种取材方法中,混合消化液培养法细胞

关 键 词:结膜上皮细胞  免疫组化  冻存复苏

The Culture and Cryopreservation of Human Conjunctival Epithelium in Vitro
Jianliang Zheng,Rong Lu,Jie Zhang,Mingkai Lin,Jianxian Lin Zhongshan Ophthalmic Center,Sun Yat-sen University of Medical Sciences. Guangzhou ,China.The Culture and Cryopreservation of Human Conjunctival Epithelium in Vitro[J].Eye Science,2000,16(2):135-138.
Authors:Jianliang Zheng  Rong Lu  Jie Zhang  Mingkai Lin  Jianxian Lin Zhongshan Ophthalmic Center  Sun Yat-sen University of Medical Sciences Guangzhou  China
Institution:Zhongshan Ophthalmic Center, Sun Yat-sen University of Medical Sciences, Guangzhou 510060, China.
Abstract:Purposes: To detect the best method of culture and preservation of human conjunctival epithelia. Methods: Human conjunctival epithelia were cultured by tissue inoculation, mechanical separation or enzyme digestion with 0. 25% trypsin. The cultured cells were identified through their morphology, growing features and immunohistochemical staining. The third and fourth passage confluent cells were frozen in liquid nitrogen, and resuscitated 30 days later. Results: Cells from tissue digested with 0.25% Trypsin grew much better than those from tissue inoculation. No cell outgrew in mechanical separating group. The cultured cells spread along the flask in polygonal shape and positive in pan-Keratin staining. Ninty percent of the cryopreserved cells were successfully resuscitated . Conclusion: Tissue digestion with trypsin is the best approach to culture human conjunctival epithelium, and the cultured cells can be cryopreserved in liquid nitrogen. Eye Science 2000; 16: 135 - 138,
Keywords:conjucntival epithelium  immunohistochemistry  cryopreservaton resuscitation
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