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将生后3～4天大鼠视网膜细胞分离,培养在人羊膜基膜(HABM)、基板蛋白(LN)和多聚鸟氨酸(PO)支持物上。分为加顶盖提取液(Te)和未加Te对照两组,以形态学和四唑盐(MTT)微量自动比色法对细胞生长进行定量测定。用神经元特异性烯醇酶(NSE)抗体作免疫细胞化学染色,以便辨认神经细胞。所得结果表明,经48小时培养,加Te的HABM、LN和PO实验组与相应的对照组比较,视网膜细胞生长更活跃。在有Te的HABM和LN上生长的细胞直径,超过10μm的比无Te的对照组多3倍。从MTT光密度值显示,加Te的实验组细胞,生长在7×10~4～1.2×10~5/孔的密度范围内,与无Te的对照组相比差异非常显著(P<0.01);尤以1×10~5细胞密度/孔的生长最为活跃。在细胞密度低于6×10~4/孔时,细胞生长不活跃。证实在一定的视网膜细胞密度范围内,Te对其生长有明显的促进作用。NSE免疫细胞化学染色证实,90％以上的视网膜分离培养细胞为阳性,其中绝大部分为小细胞,其直径为5～6μm,大细胞直径在10μm以上的约占0.1％。
Rat retinal cells were isolated 3 to 4 days after birth and cultured on human amniotic membrane (HABM), substrate protein (LN) and polyornithine (PO) support. Divided into plus cap extract (Te) and no Te control group, morphological and tetrazolium (MTT) trace auto-colorimetric method for quantitative determination of cell growth. Neuronal-specific enolase (NSE) antibodies were used for immunocytochemical staining in order to identify neuronal cells. The results showed that, after 48 hours of culture, plus Te HABM, LN and PO experimental group compared with the corresponding control group, retinal cell growth more active. Cell diameters growing on HABM and LN with Te were 3-fold more than 10 m in the Te-free control group. From the MTT optical density values, the cells in the experimental group with Te were grown in the density range of 7 × 10 ~ 4 ~ 1.2 × 10 ~ 5 / well, which was significantly different from the control group without Te (P <0.01) ; Especially in 1 × 10 ~ 5 cell density / hole the most active growth. At a cell density of less than 6 × 10 -4 / well, cell growth is not active. Confirmed that in a certain range of retinal cell density, Te has a significant role in promoting its growth. NSE immunocytochemical staining confirmed that more than 90% of the retina cultured cells were positive, most of them small cells, the diameter of 5 ~ 6μm, large cell diameter of more than 10μm accounted for about 0.1%.