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目的研究分离、培养人脑胶质瘤细胞方法。方法手术获得切除胶质瘤新鲜标本30例,酶消化法分离培养获得人脑胶质瘤细胞,利用含10%胎牛血清的DMEM培养液进行体外原代培养及传代,观察肿瘤细胞生长特点。运用冻存液冻存培养的胶质瘤细胞,分别于冻存后1、4、8、12、16、20、24周复苏,计数复苏后活细胞比例,进行再培养并观察肿瘤细胞生存情况。结果从脑胶质瘤标本分离出的胶质瘤细胞可进行原代培养及传代。冻存1、4、8、12、16、20、24周后脑胶质瘤细胞复苏的成活比例分别为54%、52%、52%、50%、48%、46%、40%。复苏后再培养的肿瘤细胞生长良好。结论通过分离培养手术切除胶质瘤标本可获得脑胶质瘤细胞株,肿瘤细胞可通过冻存的方式进行保存,冻存后复苏的细胞可再培养。
Objective To study the methods of isolation and culture of human glioma cells. Methods Thirty new cases of glioma were obtained by surgery. Human glioma cells were isolated and cultured by enzyme digestion. Primary cultured and passaged DMEM medium containing 10% fetal bovine serum was used to observe the growth characteristics of tumor cells. The frozen glioma cells were cryopreserved, resuscitated at 1, 4, 8, 12, 16, 20 and 24 weeks after cryopreservation respectively. The proportion of viable cells after resuscitation was counted, and the survival of tumor cells was observed . Results Glioma cells isolated from glioma specimens can be primary cultured and passaged. The survival rates of glioma cell resuscitation were 54%, 52%, 52%, 50%, 48%, 46% and 40% respectively after cryopreservation 1, 4, 8, 12, 16, 20 and 24 weeks. Tumor cells cultured after resuscitation grow well. Conclusion Glioma cell lines can be obtained by resection of glioma specimens. The tumor cells can be preserved by cryopreservation, and the cells recovered after cryopreservation can be re-cultured.