The study of Corneal epithelial Cell Transplantation

我们试图开发一种专为兔角膜上皮细胞设计的高贵的无血清细胞培养液。首先，当在所测试的基础培养液中考察Ca~(2+)浓度的影响时，浓度为0.03 mM的Ca~(2+)~(2+)在培养的角膜上皮细胞中表现出最大的生长。因此，利用此钙离子浓度，我们比较了MEM、MCISBI53、F-10、MCDB153和F-10(1：1)、MCDB153和MEM(1：1)的基本培养基的生长能力。结果表明，由MCDBI53和F-10(1：1)组成的培养基细胞生长最好。第三，在此基础上添加HEGF、氢化可的松、胰岛素、BPE(牛脑垂体提取物)，成功地将-70℃冷冻的原代培养细胞传至第15代。本文报道的培养兔角膜上皮细胞的基础培养液远远优于现有的基础培养液。我们尝试在IV型胶原片上建立体外兔角膜上皮细胞层。为探讨体内结膜上皮细胞在特定环境下是否发生生物变化，采用免疫组织化学方法检测了角膜中央基质袋内植入的结膜上皮细胞的生物学特性。用于角膜上皮细胞。结果提示，结膜上皮的生物学特性受外界环境的影响，与角膜上皮相似。

We tried to develop a noble serum-free cell culture medium specially designed for rabbit corneal epithelium. First, when the effect of Ca^<2+> concentration was investigated in a basal medium tested, the Ca^<2+> concentrate : Lon of 0.03mM showed maxiTmnn growth in cultured corneal epithelial cells. Therefore, using this Ca^<2+> concentration, we compared the growth ability of basal medium among MEM, MCISBI53, F-10, MCDB153 and F-10(1 : 1), MCDB153 and MEM(1 : 1). The results indicated that the medium consisting of MCDBI53 and F-10(1 : 1)showed the highest cell growth. Third, using this basal medium supplemented with HEGF, hydrocortisone, insulin, BPE(bovine pituitary extract), primary cultured cells frozen at -70 C were successfully subcultured up to the 15th-generation. The medium reported here is presixmed to be far superior to any other available basal medium for culturing rabbit corneal epithelium.We tried to establish in vitro rabbit corneal epithelial cell layer on type IV collagen sheet. Since two to three layers of cuboidal epithelial cells were successfully made one week after initial culture, this epithelial sheet can be transplanted into in vivo ocular surface.We investigated whether biological alteration truly occurs in in vivo conjunctival epithelium in a certain environment, the biological characteristics of conjunctival epithelium implanted in a central corneal stromal pocket were immunohistochemically examined using 64Kd keratin expression, a specific differentiation marker. for corneal epithelium. The results suggest that the biological characteristics of conjunctival epithelium can be influenced by the external environment, becoming similar to those of corneal epithelium.