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目的获得高纯度的NF1(Ⅰ型神经纤维瘤病)神经纤维瘤中雪旺细胞。方法将手术室取来的NF1神经纤维瘤组织剪碎,用Ⅰ型胶原酶消化,普通培养液吹打分散差数贴壁后接种于培养瓶,细胞贴壁后换用含Geneticin的培养液培养3 d,换用普通培养液继续培养,待细胞汇合时结合低浓度酶快速消化法和差数贴壁法对雪旺细胞进行纯化,传代培养;继用S-100蛋白进行免疫组化染色,鉴定培养的雪旺细胞。结果应用含Geneticin的培养液培养雪旺细胞,结合酶快速消化法和差数贴壁法,成纤维细胞基本消失,雪旺细胞活力良好,可进行传代培养,经S-100蛋白免疫组化染色鉴定培养的细胞基本为染色阳性的雪旺细胞。结论应用含有Geneticin的培养液培养雪旺细胞,结合酶快速消化法和差数贴壁法,能够有效地去除成纤维细胞,获得纯度高、活力良好的雪旺细胞。
Objective To obtain Schwann cells in high purity NF1 (type Ⅰ neurofibromatosis) neurofibromas. Methods The NF1 neurofibroma tissue taken from the operating room was cut, digested with type Ⅰ collagenase, dispersed in ordinary culture medium and then spread on the culture flask. The cells were fixed in culture medium containing Geneticin d, to continue with ordinary culture medium, until the cell confluency combined with low concentrations of enzyme digestion and poor adhesion method of Schwann cells purified, subculture; following the S-100 protein immunohistochemical staining, identification Cultured Schwann cells. Results The Schwann cells were cultured with the culture medium containing Geneticin, fibroblasts were almost disappeared by combined enzymatic digestion and differential adhesion method, and the Schwann cells were viable and subcultured. S-100 protein immunohistochemical staining The cultured cells were identified as stained positive Schwann cells. CONCLUSION: Schwann cells cultured in a medium containing Geneticin can be used to remove fibroblasts effectively by combining enzymatic digestion and differential adherent method, and to obtain pure and viable Schwann cells.