Therefore, further study of EGF in the tissue engineering has a scientific significance

Therefore, further study of EGF in the tissue engineering has a scientific significance. In view of above evidence, we aimed to explore the role of EGF produced by HaCaT cells in the proliferation, invasion, migration and transdifferentiation of ADSCs into epidermal cell phenotypes. expression of EGF was up or down regulated constantly in HaCaT cell collection after transfection. EGF overexpression upregulated the proliferation, migration and invasion rates of ADSCs, and EGF expression regulated the expression of cytokeratin-19 (CK19) and integrin-as well. Conclusions EGF could be served as a stimulus to promote the proliferation, migration, and invasion as well as the transdifferentiation into epidermal stem cell immunophenotyping of ADSCs. The results showed that EGF experienced a encouraging effect on the repair of skin wound. (4, 5). The amount of ADSCs in the same tissue is much larger than that in the bone marrow mesenchymal stem cells, and ADSCs, which have good self-proliferation and multi-lineage differentiation potentials, can differentiate into tissue cells such as adipocytes, osteoblasts, hepatocytes and endothelial cells (4, 5). Brzoska et al. (6) reported that all-trans retinoic acid (ATRA) induced the differentiation of adipose-derived stem cells into epidermal cells, indicating that adipose-derived stem cells have the ability to differentiate into epidermal cells across the germ layer. Yao et al. (7) revealed that this transplantation of ADSCs could promote the healing of skin deep partial-thickness scald wound of rabbit. Thus, the usage of ADSCs in the repair of wound skin shows a encouraging effect. Epidermal growth factor (EGF) is usually a type of polypeptide, which is composed of 53 amino acids and promoting mitosis (8). After hydrolysis, it exerts its biological activity on and participates in the progress of skin proliferation, differentiation, apoptosis and carcinogenesis (9). Previous studies had shown that EGF could induce the early development of teeth and eyelids in mice inhibit the secretion of gastric acid, as well as promote the growth of epidermis and the keratinization process (8). In addition, experts also reported that ADSCs treated with EGF by three-dimensional culturing method would differentiate to an epithelial phenotype (10, 11). Therefore, further study of EGF in the tissue engineering has a scientific significance. In view of above evidence, we aimed to explore the role of EGF produced by HaCaT cells in the proliferation, invasion, migration and transdifferentiation of ADSCs into epidermal cell phenotypes. Our study provides supporting evidences for ADSCs to be used as Ki8751 ideal seed cells in tissue engineering. Materials and Methods The extraction of ADSCs and cell culturing The protocol of this study was approved by the ethics table of Sun Yat-sen Memorial Hospital, Sun Yat-sen University or college (approval number: SY2017010745). Adipose tissue was collected from a female individual who aged 29 years old. The patient signed the knowledgeable consents before the study has been conducted. Under sterile conditions, about 10 g of the remaining abdominal subcutaneous adipose tissue of the patient who required the plastic surgery was sent to the laboratory within 1 h. The tissue was first immersed in PBS (Solarbio Life Sciences, Beijing, China) Ki8751 made up of penicillin (300 U/ml) and streptomycin (300 for 30 min following the instructions Rabbit polyclonal to ADNP2 of manufacturer. The supernatant Ki8751 of cells was gathered and stored at ?80. The concentration of total protein was determined by the PierceTM BCA Protein Assay Kit (Thermo Fisher, Waltham, USA). 25 as well To investigate whether the expression of EGF could regulate the proliferation of ADSCs and promote the transdifferentiation of ADSCs into epithelial stem cell types, we measured the cell viability of ADSCs by MTT, decided the protein levels of EGF in the bottom chamber and assessed the expressions of CK19 and integrin-in ADSCs. On one hand, the results from the co-cultured ADSCs and HaCaT cells showed that this cell viability in HaCaT-EGF group was higher than that in HaCaT-Mock group, however, no significance between HaCaT-NC and HaCaT-siEGF groups (Fig. 3A, *p<0.05) was observed. Besides, the protein level.