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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%。
Objective: To investigate the ratio of Schwann cells derived from neural crest in the sciatic nerve. Methods: Wnt1-Cre +/- and Rosa-EGFP +/- mice were crossed to obtain Wnt1 / EGFP mice. EGFR protein was expressed in all cells of neural crest. Take the sciatic nerve, digested and purified to obtain Schwann cells. Anti-GFP immunofluorescence staining and flow cytometry were performed. Results: According to morphological observation of Schwann cells in P1 generation, the purity was about 60%. Anti-GFP immunofluorescence staining showed that not all Schwann cells were positive. Purity of P3 generation Schwann cells is about 99%, and flow cytometry analysis shows a GFP-positive rate of about 65%. CONCLUSION: Schwann cells in the sciatic nerve of mice are cultured and purified in vitro. The proportion of Schwann cells originating from neural ridge to the total number of Schwann cells is about 65%.