人角膜缘干细胞提取方法的比较性研究  

A comparative study on procedures to extract limbal stem cells from human corneas

作　　者：齐飞[1] 于佳欣[2] 李若溪[1] 

机构地区：[1]沈阳市第四人民医院,辽宁沈阳110031 [2]辽宁中医药大学

出　　处：《中国伤残医学》2015年第8期31-33,共3页Chinese Journal of Trauma and Disability Medicine

摘　　要：目的：比较组织块培养法、酶消化法及平板离心法获取角膜缘干细胞的不同。方法：取得人组织，分别应用组织块培养法、酶消化法及平板离心法获得角膜缘细胞并进行原代细胞培养，对获得的细胞通过免疫组化方法研究其p63、ABCG2及K3表达情况，以比较上述3种方法的不同之处。结果：3种方法均可获取大量阳性表达p63、ABCG2及阴性表达K3的细胞，符合角膜缘干细胞特点，但其中组织块培养法中阴性表达p63的细胞略多，并偶见阳性表达K3的细胞；另外，在提取细胞的实验操作方面，组织块培养法耗时约7天，酶消化法耗时约1天，平板离心法耗时约2天；在获得细胞量方面，1枚人角膜缘材料酶消化法可获得的细胞数量在此3种方法中通常最少。结论：平板离心法在目前常用的人角膜缘干细胞提取方法中可能是最具优越性的。Objective：To study the difference among the methods to extract limbal stem cells from human corneas.Methods：Limbal cells were extracted from donated human corneas with 3 different procedures：explant culture （G1）, single cell suspension （G2） and un-coated centrifugation （G3）.The expression of p63, ABCG2 and K3 of the primary cultures of the cells were studied.Results：All 3 pro-cedures produced good quantity of cells being p63 positive, ABCG2 positive and K3 negative.But some p63 negative cells were seen in G1, and a small amount of K3 positive cells were seen in G1 and G2.The procedure of cell extraction took about 7 days in G1, and about 1 day in G2 and in G3.And the amount of cells extracted from 1 cornea in G1 was often the least among the 3 procedures.Conclusion：Uncoated centrifugation may be the best among the 3 procedures.

关 键 词：角膜缘干细胞 组织块培养法 酶消化法 平板离心法 

分 类 号：R779.65[医药卫生—眼科]