神经脊来源许旺细胞的体外培养研究

目的:探讨坐骨神经中神经脊来源的许旺细胞所占的比率。方法:将Wnt1-Cre+/-与Rosa-EGFP+/-小鼠杂交,获取Wnt1/EGFP小鼠,其所有起源于神经脊的细胞都有EGFP蛋白表达。取其坐骨神经,经过消化分离纯化,获得许旺细胞。进行抗GFP免疫荧光染色和流式分析的检测。结果:根据P1代许旺细胞的形态学观察,其纯度约为60%。抗GFP免疫荧光染色显示,并非所有的许旺细胞均呈阳性。P3代许旺细胞的纯度约为99%,流式细胞术分析显示约65%左右的GFP阳性率。结论:小鼠坐骨神经中的许旺细胞在体外培养提纯后,神经脊起源的许旺细胞占总许旺细胞的比例约为65%。

Research of Schwann Cell Derived from Nerve Crest in Vitro

Objective:To investigate the percentage of Schwann cells arose from neural crest in a rate model.Methods:Wnt1-Cre+/-with Rosa-EGFP+/-mice were mated to get the Wnt1/EGFP mice, whose cells were originated from neural crest with GFP labelling among their prognance. Then the sciatic nerve were taken out and digested. Schwann cells were harvested and cultivated. The purified Schwann cells were used to do immunofluoresence staining and flow cytometry analysis.Results:In P1 passage of Schwann cells, the purity was 60% and not all Schwann cells were positively labelled in anti-GFP immunofluoresence. In P3 passage of Schwann cells, the purity was 99%, and about 65% of them originated from neural crest in flow cytometry analysis.Conclusion:About 65% of Schwann cells in vitro arose fromneural crest in sciatic nerves in mice.