人、兔结膜上皮细胞的原代培养

目的探索结膜上皮细胞体外培养的最佳方法，研究其生长特性，为毒理实验及体外结膜上皮移植提供基础。方法分别用3种培养方法：(1)组织块培养法；(2)混合肖化法；(3)组织块消化后贴壁法来培养人、兔的结膜上皮细胞，观察细胞状态、不同部位的长生特性等，并尝试在羊膜、饲细胞层上培养建系。结果3种培养方法中，组织块培养法能获得较纯的结膜上皮细胞但生长缓慢，消化法能获得大量细胞，生长较快，但常常混有其它杂细胞，组织块消化后贴壁法获得细胞量较少。观察到来自兔、睑结膜及穹隆部结膜上皮的细胞生长速度较快，细胞量多，来自球结膜的细胞生长较慢。应用间接免疫荧光染色，人结膜上皮细胞K13染色阳性。结论组织块培养法及混合消化法皆能培养结膜上皮细胞，各有利弊。

Primary Culture of Conjunctival Epithelial Cells of Human and Rabbit in Vitro

Objective To study the best culture method an d growth characteristics of conjunctival epithelial cells of human and rabbits in vitro and to provide a model as a base for toxicity experiment and clinical tran splantation.Methods 3 culture methods were used,namely:(1) tissue ino culation;(2) mixed enzyme digestion;(3) tissue adhere after digestion.The cell m orphology and in vitro growth features at different locations would be observed and the cell line was tried to be set up in AM (amniotic membrane) or 3TS 3 cell s.Results Pure conjunctival epithelial cells could be obtai ned by tissue inoculation method but they grew slowly.A large amount of cells co uld be obtained by mixed enzyme digestion method and they grew quickly.Cell numb ers were limited by method 3.Cells from palpebral and fornix grew more quickly t han those from bulbar.Cells could be identified by CK 13 by using indirect immun fluorescence stainig.Conclusion The method 1 and method 2 all can be used to c ulure conjunctival epithelial cells of human or rabbit in vitro,but each has its own advantage or disadvantage.