摘要
目的:通过对比并改良常见的原代(CAFs)细胞培养方法,建立一种成功率较高的宫颈癌相关成纤维细胞(CAFs)原代培养的方法。方法:取确诊为宫颈癌患者的活检或手术标本,通过对消化时间、重悬液量及培养基血清浓度进行改良原代细胞培养,观察细胞生长状况,并比较其与传统组织块贴壁法和酶消化法的细胞生长率。运用时间差酶消化及反复贴壁法进行分离纯化后,用免疫荧光的方法对宫颈癌相关成纤维细胞的纯度进行鉴定。结果:改良培养法消化时间为30min,血清浓度为20%时成功率最高(66.67%)。反复运用时间差酶消化及反复贴壁法可以得出纯度较高的宫颈癌相关成纤维细胞。结论:本研究成功建立了一种成功率较高的宫颈癌相关成纤维细胞改良培养方法,为后期探讨CAFs在肿瘤形成、进展中的作用打好实验基础。
Objective:To establish a primary culture method of cervical cancer associated fibroblasts(CAFs)with high success rate by comparing and modifying common primary cell culture methods.Methods:Cervical cancer specimens were obtained from patients underwent biopsy or operation,the primary culture was performed which differred from traditional method in digestion time,suspending liquid volume and serum concentration in culture medium,and cell growth condition was observed.Then,cell growth rate of traditional tissue adherent method and enzymatic digestion method was compared.In addition,after isolation and purification by time difference enzyme digestion and repeated adherent method,the purity of cervical cancer associated fibroblasts was identified by immunofluorescence.Results:The modified primary culture with digestion time of 30 minutes and serum concentration of 20% was proved to be a method with higher success rate(66.67%).Immunofluorescence showed that the high purity of cervical cancer associated fibroblasts could be obtained by repeating use of time difference enzyme digestion and repeatedadherence method.Conclusion:A modified primary culture method of cervical cancer associated fibroblasts may be successfully established in this study,which lay a foundation for further revealing the mechanisms of CAFs in tumor formation and progression.
出处
《武汉大学学报（医学版）》
CAS
2017年第2期193-197,共5页
Medical Journal of Wuhan University
关键词
原代培养
宫颈癌
成纤维细胞
改良法
Primary Culture
Cervical Cancer
Cancer Associated Fibroblasts
Modified Eethod
