STUDY ON THE PROTECTIVE SOLUTIONS OF hRPE CELLS AFTER BREAKING AWAY CULTURE CONDITION IN VITRO
- VernacularTitle:视网膜色素上皮细胞脱离体外培养条件后细胞保存液的研究
- Author:
Shucong MENG
;
Jun ZHANG
;
Xiaoyan ZHANG
;
Aili LU
- Publication Type:Journal Article
- Keywords:
Human retinal pigment epithelium (hRPE);
Cell apoptosis;
Cell protective solutions;
Fluorescence activated cell sorter
- From:
Acta Anatomica Sinica
2002;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the best protective solution that could maintain the cell activity and prolong the survival time of the human retinal pigment epithelium (hRPE) cells after break away in vitro culture case. Methods hRPE cells (stored in our laboratory) culture in DMEM/F12, added 10% FBS, EGF 20?g/L and bFGF 20?g/L, which had been cultured for more than five generations. When these culture hRPE cells had been confluenced more than 80%, the hRPE cells were harvested and placed into six kinds of protective solutions (balanced salt solution, Quinn's advantage medium, physiological salt solution, 18-amino acid solution, 5% glucose solution, artificial cerebrospinal fluid), then the cells were stained with the Annexin V /FITC, and the numbers of cell death and cell apoptosis were detected with the fluorescence activated cell sorter (FACS). The hRPE cells were in six kinds of protective solutions and were investigated at three different times from 2 hours to 8 hours. Results The result presented as follows: At the room temperature, the hRPE cells in all six kinds of protective solutions occurred necrosis within 30 minutes. To investigate the state at 4℃, in a duration of 2 hours, the apoptosis rate in the group of balanced salt solution was lowest (0.9%); As compared with the group of balanced salt solution, the apoptosis rate of artificial cerebrospinal presented a great difference (P
