Primary isolation, culture and biological property analysis of breast cancer-associated fibroblast

VernacularTitle:乳腺癌中肿瘤相关成纤维细胞原代分离、培养及相关生物学特性分析
Author: Chunmin ZHANG ; Bianqin GUO ; Lixiang WU
Keywords: Breast neoplasms; Fibroblasts; Cell culture techniques; Neoplasm invasiveness; Cell movement
From: Cancer Research and Clinic 2017;29(8):510-514
CountryChina
Language:Chinese
Abstract: Objective To explore a stable and feasible method for the primary culture of breast cancer-associated fibroblast (CAF), and to offer the theoretical basis for the further studies about the role of CAF in the processing of breast cancer by analyzing the biological property of CAF. Methods CAF was primary cultured with tissue block enzymolytic method. Flow cytometry (FCM) was used to detect the cell cycle. The expression of Vimentin, α-SMA and Ki-67 were detected by cell immunochemistry method. Transwell assay was used to detect the invasion and migration capacity of CAF. Results The CAF was successfullyculturedbyconditionoptimization.ImmunohistochemicalresultsshowedthatVimentin, α-SMA and Ki-67 were highly expressed in CAF.Comparedwithfibroblastsfrombreastcancersidestissues,the proliferating ability and the invasion and migration capacities of CAF were enhanced (number of invasive cells: 79.7 ± 3.5 vs. 66.3 ± 1.5, number of migrated cells: 242.3 ± 3.1 vs. 218.3 ± 2.1, both P<0.05). Conclusion CAF cultured from breast cancer has better proliferating ability and strong invasion and migration capacities.