SD大鼠视网膜小胶质细胞原代培养方法的改良

目的对经典的原代视网膜小胶质细胞体外纯化培养方法进行简单的改良以提高细胞产量。方法在视网膜小胶质细胞原代培养过程中以McCarthy等的经典方法为基础,寻找适当的初始种植密度,并在初次振荡分离后继续培养以获得多次产出,使用免疫细胞化学染色方法进行小胶质细胞纯度鉴定。结果视网膜小胶质细胞原代培养最宜初始种植密度为1×106/mL,多次分离纯化获得的视网膜小胶质细胞均达到97%以上的纯度。结论视网膜小胶质细胞原代培养过程中,以适当的初始种植密度和多次振荡分离方法可在保证有效纯度的基础上提高细胞产量。

Modification of the Isolation and Purification of Rat Retinal Microglia

Objective To make a simple modification of standard culture protocol for retinal microglia in order to increase the cell harvest.Methods Based on the standard microglia culture protocol developed by McCarthy,A mixed glial culture with different seeding density was prepared from rat retina.After isolating microglia from mixed cultures for the first time,fresh media were added to the cultures for additional isolation procedures.Immunocytochemical staining was used to determine the purity of harvested retinal microglia from multiple yields.Results The most suitable seeding density was 1×10~6/mL.The purity of retinal microglia from multiple isolations was higher than 97%.Conclusion With appropriate seeding density and repetitive isolation,this modified protocol can be used to increase overall harvest without scarifying purity.