Mixed Cells Of Hair Follicles Induced Exploration And Study Of The Hair Follicle Renewal

Object: Hair follicle outer root sheath cells (HHFORSC) and human hair follicle dermal papilla cells (HHDPC) were bought from ScienCell company and isolated and cultured by ourselves to obtain sufficient cells. HHDPC, HHFORSC were seeded into 3D alginate scaffold to construct three-dimensional model of hair follicle in vitro. Then implanted the model into subcutaneous tissue of SD rats in order to induced the formation of hair follicle structure . At the same time ,to explore the biological characteristics of the hair follicle and the role of those two kind of cells in the formation of follicle.Method: Human Hair Follicle Outer Root Sheath Cells(HHFORSC),Human Hair Dermal Papilla Cells(HHDPC) were bought from ScienCell company, then recoveried and cultured. At the same time, using enzymatic digestion and tissue culture to get HHDPC and HHFORSC. Comparing the growth characteristics of HHORSC and HHDPC from two different ways after cultured to get sufficient cells. Both cells mixed and seeded into alginate 3D cell scaffold to construct hair follicles in vitro culture for 7 weeks. Observing the situation of follicle formation by light microscope. Meanwhile, transplanted the three-dimensional model into the subcutaneous tissue of SD rats . After 8 weeks, the transplant sites from SD rats were harvested and observed by HE,immunohistochemistry and electron microscopy.Results: There were not significant difference in biologic characteristics between the HHDPC and HHFORSC cells bought from ScienCell company and got by enzymatic digestion and tissue culture. Hair follicle cells scattered evenly in three-dimensional scaffold were seen under light microscope after culture in vitro for 7 weeks , but keratinized material and hair follicles can't be seen. SD rat were harvested after 8 weeks, slices from the transplant site showed cells gathered and formed cell aggregate, look like hair follicles-like structure under light microscope by HE staining. CK14, CK15 and vimentin staining of the cell aggregate showed positive. Cells scatted into scaffold by electron microscopy observation. Conclusions: HHDPC and HHFORSC cells got by enzymatic digestion and tissue culture kept a good growth characteristics. There was not follicle-like structures be seen in three-dimensional reconstruction of hair follicle in vitro . Slices from the transplant site showed cells gathered to form hair follicles-like structure by HE staining observation. CK14, CK15 and vimentin stainong of cell aggregate showed positive. It indicated that hair follicles-like structure could be induced in vitro culture, HHDPC keep the inducing function in hair follicle, HHFORSC keep the activity of hair follicle stem cell. This reconstruction method could provide a direction for hair follicle reconstruction.