In Vitro Induced Differentiation Of Human Breast Adipose-derived Stem Cells Into Breast Epithelial-like Cells

Part Ⅰ Isolation, culture and identification of adipose derived stem cells from human hypertrophy breast adipose tissueObjective:To establish a method for isolating human breast adipose derived stem cells(hbASCs)from resected human hypertrophy breast adipose tissue. Methods:hbASCs were isolated, cultured, and expanded from human hypertrophy breast adipose tissue. Immunofluorescent staining of specific molecules, FACS and multilineage differentiation induction were used to characterize the obtained hbASCs. Results: hbASCs obtained in this study had the characteristics of mesenchymal stem cells and expressed specific molecules; they also possessed a multilineage differentiation potential. Conclusion:hbASCs can be isolated from human hypertrophy breast adipose tissue,which provides a novel and abundant seeding cells for tissue engineering. Part II In vitro induction of human breast adipose-derived stem cells (hbASCs) into breast epithelial-like cells by co-culturingObjective:To explore the feasibility of the trans-diferentiation of human breast adipose-derived stem cells (hbASCs) into breast epithelial-like cells after co-culturing in Transwell in vitro. Methods: Experimental study. The third passage hbASC and the HBL-100cell line were co-cultured in a Transwell culture system for15days. The human MhbASCs were observed and identified by inverted phase contrast microscope and transmission electron microscopy, and immunocytochemistry staining in the induced and control groups. Results:Both the third passage hbASCs and the HBL-100cell line cells could adhere and grow rapidly after co-cuhure in the Transwell. After co-culture for15days, the morphology of some induced hbASCs changed into epithelial-like cells. Some induced hbASCs showed positive of CK18,CK19by immunocytochemistry staining, and typical epithelium cells with microvilli, desmosomes and tonofilaments that were observed under TEM. Conclusion:The data suggests that hbASCs may have the potential to transdiferentiate into human breast epithelial-like cells after co-culturing in Transwell in vitro. Part ⅢEGF promoted differentiation of hbASCs into human breast epithelial-like cells by co-culturing in vitro.Objective:To investigate the feasibility for promoting the trans-differentiation of human breast adipose-derived stem cells (hbASCs) into breast epithelial-like cells by the epidermal growth factor(EGF) after co-culturing in vitro. Methods:Experimental study. hbASCs were at passage3. Two experimental groups were established based on the following criteria of culture medium:hbASCs+CM (HBL-100)+lOug/L EGF(group1), hbASCs+CM (HBL-100)(group2). The induced cell were observed by inverted phase contrast microscope. After15days, the cell were collected for immunocytochemistry, FACS and QRT-PCR. The results were analyzed with the matched t test. Results:After co-culture, the morphology of some induced hbASCs in both groups changed into epithelial-like cells, some induced hbASCs showed positive of CK18,CK19by immunocytochemistry staining. The expression of CK18 was examined by QRT-PCR. The optical density value (683.000±58.014) and the expression of CK18(3.5350±0.3737) differences between group1and group2were statistically significant(P<0.05). Conclusion:The trans-differentiation of human breast adipose-derived stem cells (hbASCs) into breast epithelial-like cells can be promoted by the epidermal growth factor(EGF) after co-culturing in vitro.