Primary isolation, culture and biological property analysis of breast cancer-associated fibroblast / 肿瘤研究与临床

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.