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人眼虹膜色素上皮细胞体外培养、冻存与复苏

The Cultivation, Freezing-state Preservation and Resuscitation of Human Iris Pigment Epithelial Cells in Vitro

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【作者】 李莉葛坚卓业鸿林明楷郑健梁

【Author】 Li Li, Jian Ge, Yehong Zhuo, Mingkai Lin, Jianliang ZhengZhongshan Opthalmic Center, Sun Yat-sen University of Medical Science, Guang zhou 510060, China

【机构】 中山医科大学眼科中心中山医科大学眼科中心 广州 510060广州 510060广州 510060

【摘要】 目的:建立人眼虹膜色素上皮细胞体外培养并对其进行冻存与复苏。方法:直接刮取虹膜色素上皮细胞组织碎屑进行培养,光镜观察生长特性及形态特点,透射电镜观察其超微结构。根据慢冻速融的细胞冻存原则,定期收集细胞进行液氮冻存,至少2个月后进行复苏。结果:人眼虹膜色素上皮细胞体外培养成功,原代细胞在光镜下呈多角形单层生长,胞浆内有丰富的色素颗粒;电镜下见胞浆富含色素、细胞器丰富、细胞膜有明显微绒毛、微丝、相临细胞之间可见桥粒连接。共冻存6批细胞,进行4次复苏实验均成功,每次复苏细胞存活为90%。结论:人眼虹膜色素上皮细胞体外培养的建立及冻存、复苏的成功,为研究某些疾病提供有利的基础。眼科学报2000;16:220~223。

【Abstract】 Purpose: To establish the culture of human iris pigment epithelial cells in vitro and study the freezing-state preservation and resuscitation.Methods: The iris pigment epithelial specimens from human eyes were cultured in vitro. Based on the cell-frozen principle, the cultured cells were collected, then frozen in liquid nitrogen and resuscitation.Results: Cultured iris pigment epithelial cells were obtained. Under the inverted light microscope, primary cells appeared multigonal and arranged in monolayer, there were abundant pigment granules in the cytoplasm and the nuclei each of which contained 1 or 2 nuce-oli wre relatively transparent. Under the transmission electron microscope, there were plenty of microvilli at the cell membrane and desmosomes present in the intercellular space. 6 groups of cells were frozen. The resuscitation exeriment was carried out for 4 times, every times being successful. All of the resuscitated rates were more than 90%. Conclusion: The human iris pigment epithelial cells were cultured in vitro successfully, they can also be frozen and resuscitated, which will be useful in studies of pathogenesis for some eye disease. Eye Science 2000; 16: 220 ~ 223.

【基金】 国家自然科学基金(39800163);国家教委博士点基金(9856);广东省各部委基金(980112)
  • 【分类号】R77
  • 【下载频次】79
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