Research on human epidermal stem cells differentiating into glandular epithelium-like cells in vitro / 重庆医学

ABSTRACT

Objective To investigate the influencing conditions of human epidermal stem cells(ESCs) differentiating into gland-like cells and to identify the induced tubular structure.Methods The ESCs were seeded onto compound polysaccharide shell dermal matrix and collagen gel,adding different concentrations of epidermal growth factor(EGF) and culturing by vertical shaking in vitro,the three dimensional culture and induced directional differentiation were performed.The means of HE staining,immunofluorescence and confocal laser scanning microscope were adopted to observe the conditions,morphology and phenotype change of ESCs directionally differentiating into glandular epithelium-like cells.Results 15-20 ng/mL EGF could induce ESCs in tissue-engineering dermis to grow into dermis and appear the gland-like structure.The HE staining in this structure showed its profile as a single layer with lacune in the middle compartment,eosinophilic cytoplasm,lightly stained.Under CLSM,by CK19 staining,the luminal structure in the middle of the cell mass was observed,while CK18 and CEA were expressed in this structure.Conclusion Under the induction of particular concentration of EGF,the in vitro cultured human ESCs seeded on the tissues engineering dermis could form into the tubular structure,which is similar to the in vivo sweat gland secretory cells in morphology and histology.