体外获得高纯度大鼠背根神经节神经元的原代培养

目的：建立一种切实可行的胚胎大鼠背根神经节神经元培养及纯化方法。方法：用显微解剖获取足够数量的胚胎大鼠背根神经节，通过胰蛋白酶+EDTA消化、交替使用NB培养基与加入了5-氟-2-脱氧尿嘧啶核苷酸/尿苷的抗有丝分裂NB培养基等方法，在体外获得纯化的背根神经节神经元，采用神经丝蛋白免疫细胞化学染色法与DAPI染核的方法鉴定并测定神经元的纯度。结果：获得的背根神经节神经元在体外生长良好，纯度可达到96%以上。结论：本实验方法可以获得大量高度纯化的大鼠背根神经节神经元。

Primary culture method to obtain high purified dorsal root ganglion neurons of rats in vitro

ObjectiveTo establish a feasible method of culture and purification of embryonic dorsal root ganglion neurons of rats.MethodsAdequate dorsal root ganglions (DRGs) were obtained by microdissection, then were dissociated with trypsin plus EDTA. The dissociated neurons were cultured with NB median and 5-fluoro-2′-deoxyuridine/uridine for purification. Anti-NF200 and DAPI staining was to examine the purification percentage of DRGn.ResultsThe isolated and cultured DRGn survived and grew in good condition in vitro, and the purified percentage of DRGn was above 96%. Conclusion The method is very useful in obtaining the massive high purified embryonic dorsal root ganglion neurons of rats.