两种配方胰酶消化法对兔角膜内皮细胞活性影响的比较

The influence of rabbit cornea endothelial cells digested using two different trypases

目的研究两种不同配方的胰蛋白酶对兔角膜内皮细胞生物活性影响。找出一种更适合于角膜内皮细胞(corneal endothelial cell,CEC)消化的方法。方法采用0.125%胰蛋白酶和0.05%胰蛋白酶-0.01%EDTA-Na2分别对CEC进行消化、培养,用台盼蓝染色法和24h细胞贴壁率观察CEC存活率及细胞活性。结果两种配方胰酶消化CEC,细胞存活率都在95%以上,无统计学差异(P>0.05)。而细胞贴壁率,胰酶消化法<30%,胰酶-EDTA-Na消化法>90%,有统计学差异(P<0.05)。结论胰酶-EDTA-Na消化法对CEC的作用较温和,活性影响小,此方法更适合CEC的消化、培养。

Objective To investigate the influence of rabbit cornea endothelial cells （CEC） digested using two different trypases and obtain a better digestion method. Methods Rabbit cornea endothelial cells were cultured using digestion with 0.125% trypase and 0.05% trypase with 0.01% ethlenediamine tetracetic acid disoduium （EDTA-Na）. CEC were stained using solution of Teypane blue and calculated the adherence rate in 24 hours, and then the survival of CEC was obtained. Results CEC digested by different trypase showed over 95% survival rate. However, a significant difference was found in adherence rate in 24 hours （P〈 0.05）. Conclusion The digestion method using trypase-EDTA-Na was better for cultivation of CEC in comparison with trypase solo.