Abstract:

Objective To construct tissue engineered urethra with enriched rabbit hair follicle stem cells, and observe its effect in repair of urethral defect. Methods Rabbit hair follicle cells were isolated and cultured by mechanical separation and enzyme digestion, and were in vitro cultured and proliferated to 3×10⁶. Integrin-α6+/cd71－ cells of <10 μm in diameter were sorted by flow cytometry, and the expression of K19, p63 and β1 integrin was detected by immunofluorescence method. Hair follicle stem cells were seeded on bladder acellular matrix scaffold, tissue engineered urethra was constructed, and the cell distribution on scaffold was observed by scanning electron microscope. Rabbit urethral defect of 2.5 cm was repaired with tissue engineered urethra, and the effect of repair was observed by urethrography and histological examination. Results Immunofluorescence detection revealed that there was positive expression of K19, p63 and β1 integrin in hair follicle stem cells. Scanning electron microscope demonstrated that cells firmly attached to the scaffold and moderately stretched, and there was matrix secretion. Histological and immunohistochemical examinations indicated there was stratified epithelium-like structure, with positive staining of AE1/AE3 in hair follicle cells. Three months after repair of urethral defect by tissue engineered urethra, urethrography illustrated the urethra was smooth and unobstructed. Conclusion Tissue engineered urethra constructed by enriched rabbit hair follicle stem cells can be used for repair of rabbit urethral defect.

Key words: hair follicle stem cell, urethra, tissue engineering