三种小胶质细胞原代培养方法的比较

Comparison of three microglia primary culture methods

目的探讨建立简捷高效原代小胶质细胞纯化培养方法。方法以小胶质细胞原代培养McCarthy经典培养方法为基础，分为常规对照组、机械分离组、胰酶消化组和利多卡因组共四组培养，并对比观察记录形态变化并计数，借助CD68及OX42抗体间接免疫荧光标记进行鉴定、计算纯度。结果三种分离方法均获得了较高纯度和产量的小胶质细胞，以利多卡因组为佳，纯度为（94．92±7．05）％，流式细胞结果显示细胞纯度约为（96．2±2．8）％。结论小胶质原代细胞培养过程中，采用利多卡因处理可显著提高细胞分离效率及纯度。

Objective To find an easy and effective method for primary culture and purification of microglia cells. Methods Based on classical microglia primary culture method of McCarthy, four groups （control group,mechanical separation group,enzyme digestion group and lidocaine separation group） were cultured. Morphologic changes and cell count were recorded and compared. Identification and appraisement of microglia cells were under the assistance of immunofluorescence mediated by CD68 and OX42 antibodies. Results All groups got high purity and quantity of microglia cells. The highest purity was obtained in lidocaine separation group [ （ 94.92 ± 7.05 ） % ]. The purity of microglia cells was （96.2 ± 2.8） % determined by flow cytometry. Conclusion In the primary culture of microglia cells, using lidocaine separation can elevate microglia cell separation efficiency and the purity significantly.