Bone marrow mesenchymal stem cells for epidermal and skin appendage regeneration / 中国组织工程研究

ABSTRACT

BACKGROUND:Burned rat serum has been reported as an inducer that can induce differentiation of mesenchymal stem cells into epidermal cells. OBJECTIVE:To induce in vitro differentiation of bone marrow mesenchymal stem cells into epidermal cells that were transplanted alone or combined with inducers for the repair of skin wound defect and epidermal reconstruction. METHODS:Under aseptic environment, rat bone marrow was harvested to culture adherent cells using low-glucose Dulbecco’s modified Eagle’s medium. Culture cells at passage 4 were confirmed as mesenchymal stem cells by flow cytometry. Bone marrow mesenchymal stem cells were induced by Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 20%burned rat serum to differentiate into epidermal cells that were identified by immunohistochemistry. Wistar rat models of ful-thickness skin wounds were prepared and divided into three groups. The 5-bromo-2-deoxyuridine-labeled autologous bone marrow mesenchymal stem cells and induced bone marrow mesenchymal stem cells were coated singly onto a rat model of burn wounds, and rat models of burn wounds with no treatment served as controls. Wound contraction and regeneration of epidermal cells and skin appendages were observed. RESULTS AND CONCLUSION:After isolation and culture of cells for 24 hours, a few of adherent cells grew as fibroblast-like cells with fusiform shape. At 16 days, cells completely covered the bottom of bottle, exhibiting a fish or reticular arrangement. After detection by flow cytometry, cells were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 20%burned rat serum, and fusiform-shaped cells gradual y became round or oval cells. Flow cytometry analysis and immunocytochemistry results showed that cells expressed keratin, which were confirmed as epidermal cells. The results show that both the bone marrow mesenchymal stem cells transplantation alone or with necessary inductor is better for skin repair than natural healing, exhibiting a faster regeneration of skin and skin appendages. Bone marrow mesenchymal stem cells are deduced preliminarily to be involved in epidermis and hair fol icle regeneration, thereby improving skin healing.