動物角膜內皮細胞培養－初報

Animal Corneal Endothelial Cell Culture Study-Priliminary Report

李弘志(Hong-Ju Lee) ； 林昌平(Chang-Ping Lin) ； 許明木(Min-Muh Sheu) ； 陳振武(Chen-Wu Chen)

《中華民國眼科醫學會會刊》 29卷2期 (1990/05) Pp. 682-688

https://doi.org/10.30048/ACTASOS.199005.0117

摘要

本文報告以猪及兔子之角膜為材料，以trypsin分離並純化其角膜內皮細胞進行細胞培養，以培養經過三代或四我的細胞進行增殖曲線及形態之分析。猪角膜內皮細胞分別以初始濃度A組：1.5×10^6cell/ml，B組：7.5×10^5cell/ml及C組：2.5×10^4cell/ml進行培養，其中A在144小時而B在培養96小時後達到融合，C組則在培養264小時時後才達到融合，其兩倍時間分別為A組：67.72小時，B組；33.02小時，C組：33.20小時。A組之兩倍時間加長可能是細胞密度過高之故。三組融合時之細胞密度並無多大差異。兔子角膜內皮細胞以5×10^4cell/ml為初始濃度，其生長至168小時時呈部份融合而生長停滯，以trypsin打散細胞後繼續生長，其二倍時間於第一階段為30.27小時，於第二階段為77.74小時。觀察細胞之形態，猪角膜內皮細胞於增殖期間呈現纖維芽細胞之形狀，於融合後則略呈規則六角形排列，如in vivo之狀態；兔子角膜內皮細胞之形態變異頗大，於初代培養中，於融合後呈紡錘體狀排列如纖維母細胞，於次代培養則可見部份如in vivo之六角形排列，部份如紡錘狀之排列。比較形態之變化於細胞培養時，猪的角膜內皮細胞較近人類之角膜內皮細胞，而兔子之角膜內皮細胞有較大的變異性。

並列摘要

The corneal endothelial cell of pigs and rabbits were isolated and cultured with cell culture technique, then their growth kinetics were studied. The pig's corneal endothelial cell concentrations: A group: 1.5×10^6cells/ml, B group: 7.5×10^5cells/ml; C groups: 2.5×10^4cells/ml. The cell in A group reached confluence after 144 hours and B group reached confluence after 96 hours, the cell in C group reached confluence after 264 hours' culutre. The doubling time of the three groups are: 67.72hrs, 33.02 hrs, 33.20hrs respectively. The cell density at confluence are without much difference in three groups. The prolongation of doubling time in A group may be due to the overcrowded initial cell density.
Rabbit's corneal endothelial cells were cultured with 5×10^4cells/ml, initial concentration. After 168hrs, the cells become partially confluent, and the cell growth had stop, after trypsinization, the cells continue to proliferate and become completly confluent. The doubling time at first stage are 30.27hrs, and second stage are 77.74hrs. The morphology of the pig corneal endothelial cells were fibroblast-like in proliferative stage but after the confluence their arrangement become hexagonal in shape which is similar to the endothelial cell in vivo. The morphology of rabbit corneal endothelial cell have much variation. In primary culture the cell become spindle in shape at confluence. In subculture, most of the cell become hexagonal shape in the central, but some of the cell still possesed spindle shape. When compared the corneal endothelial cells in this two species, the pig corneal endothelial cells are more resembling the human corneal endothelial cell than that of rabbit.

並列關鍵字

Corneal Endothelium ； Cell culture ； Doubling time ； growth curve