兔角膜上皮细胞体外原代培养方法的研究

Culture of rabbit primary corneal epithelial cells

目的:建立体外原代培养兔角膜上皮细胞简单经济实用的方法,并观察其体外生长的生物学特性。方法:采用消化法、组织块培养法培养兔角膜上皮细胞并进行传代培养,分别测定细胞分裂指数及细胞贴壁率,MTT比色法测定细胞生长曲线。结果:两种方法培养的细胞在原代、第1代形态相似,消化法培养的细胞代数维持低于组织块法;对数生长期时,组织块培养法的细胞具有更好的活力且分裂旺盛(P<0.05);在16 h后,消化法培养的细胞贴壁率低于组织块培养法(P<0.05)。结论:选择组织块培养法培养兔角膜上皮细胞可获得状态良好的连续传代扩增的细胞,且具有经济实用性;角膜上皮细胞符合一般有限细胞系的生长规律。

Objective To investigate the primary culture technique in vitro, which is convenient, economic and pragmatic, for corneal epithelial cells from rabbit＇s corneal materials. Methods The primary corneal epithelial cells were obtained by either digestion or tissue culture. The morphological characteristics of culture cells of both methods were observed by inverted phase contrast microscope at various time periods respectively. And the auxodrome （ by MTT reduction assay ）, mitotic index and adherence rate of cells were investigated as well. Results Morphological characteristics of cultured corneal epithelial cells from both methods were resembled in primary and first passage. Comparing to digestion, the cells cultured from tissue culture possessed better proliferative ability in logarithmic phase （ P 〈 0.05 ） , and the adherence rate of cells from tissue culture was higher from the 16th hour （ P 〈 0.05 ）. Conclusion Tissue culture is a simple,convenient and practicable method for culturing corneal epithelial cells which could be serially subcultivated.