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Making of Sheet of Culture Corneal Epithelium and Keratocyte by Thermal-response Cell Culture Dish.

热响应细胞培养皿制作角膜上皮和角膜细胞培养片。

基本信息

批准号：
14571688
负责人：
SHINOZAKI Kazurmi
金额：
$ 1.73万
依托单位：
Tokyo Women's Medical University, School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

The biomedical engineering research at our University has developed the thermal-response culture dish. The technique allows the collection of cell culture on a sheet without trypsins. With this method, we reproduced a corneal epithelium and a keratocyte layer with corneal epithelial cells and keratocyte cells. The sheets of corneal epithelial cells have so far been made with 3T3 cells as the feeder. However, in clinical applications, cultures from homogenous cells or serum are more preferable. We compared the reproduced corneal epithelium layers of white rabbits and of humans using : 1)keratocytes as the feeders as compared with the standard 3T3 cells ; 2)respective homogenous serum added to culture solutions as compared with the standard FBS ; and 3)different amount of growth factors. Similarly, reproduced keratocyte layers of white rabbits and humans using respective homogenous serum added to culture solutions as compared with the standard FBS were examined. The cell sheets made were examined under an optical microscope and a transmission electron microscope. The reproduced corneal epithelial cell sheets and the keratocyte sheets were transplanted to a white rabbit. The procedure allowed us to collect a reproduced keratocyte sheet similar to the collection of a 3T3 cell sheet. Although a cell layer could be reproduced successfully, a clear histological division of epithelium and other corneal layers as seen in a real cornea was not found. Serum reproduction was possible in both FBS and other homogenous serums. Whether a different amount of growth factor added to the culture solution could make a difference was unclear. At present, a reproduced epithelium layer and a keratocyte layer could be transplanted to the cornea of a white rabbit. However whether a reproduced keratocyte layer can substitute the original substance is unclear.

我校生物医学工程研究所研制了热反应培养皿。该技术允许将细胞培养物收集在没有胰蛋白酶的片材上。用这种方法，我们复制了角膜上皮和角膜上皮细胞和角膜细胞层。到目前为止，角膜上皮细胞片是用3 T3细胞作为饲养层制成的。然而，在临床应用中，更优选来自同质细胞或血清的培养物。我们比较了复制的白色兔和人的角膜上皮层，使用：1）与标准3 T3细胞相比，角膜细胞作为饲养细胞; 2）与标准FBS相比，添加到培养溶液中的各自的均质血清;和3）不同量的生长因子。同样地，与标准FBS相比，使用加入到培养液中的各自的均质血清，检查了白色兔和人的再生角膜细胞层。在光学显微镜和透射电子显微镜下检查所制备的细胞片。将复制的角膜上皮细胞片和角膜细胞片移植到白色兔中。该程序使我们能够收集类似于3 T3细胞片的复制角膜细胞片。虽然细胞层可以成功地复制，但没有发现在真实的角膜中看到的上皮和其他角膜层的清晰组织学分裂。在培养液中加入不同数量的生长因子是否会产生影响尚不清楚。目前，可以将再生的上皮层和角膜细胞层移植到白色兔的角膜中。然而，再生的角膜基质细胞层是否可以替代原始物质尚不清楚。