兔膀胱脱细胞基质负载大鼠毛囊干细胞的体外培养

In vitro culture of rabbit bladder acellular matrix carrying rat hair follicle stem cells

背景:组织工程学的兴起为泌尿系组织或器官的修复和重建开辟了新的途径,膀胱脱细胞基质是较好的泌尿系组织工程修复的替代材料。目的:构建毛囊干细胞与异种膀胱脱细胞基质为支架的细胞支架复合物,体外培养观察细胞在支架上的生长状态。方法:制备新西兰兔膀胱脱细胞基质,通过扫描电镜和Masson染色检测材料脱细胞效果,采用二次沉淀法将第3代毛囊干细胞静态接种于膀胱脱细胞基质表面,倒置显微镜下观察细胞生长状态,并绘制细胞生长曲线,组织学检测和扫描电镜观察细胞在材料表面的生长状况。结果与结论:制备的膀胱脱细胞基质为白色半透明膜状,扫描电镜下观察为纤维网状结构,未见细胞残留。Masson染色提示膀胱脱细胞基质为胶原结构,无明显细胞残留。细胞材料体外复合培养48 h后倒置显微镜下观察膀胱脱细胞基质周围毛囊干细胞生长状态良好,1周后扫描电镜下观察毛囊干细胞伸展、黏附于支架表面。结果可见毛囊干细胞与膀胱脱细胞基质体外培养具有良好的生物相容性。

BACKGROUND： The rise of tissue engineering has opened up new ways for tissue repair and reconstruction of the urinary tract, and the bladder acellular matrix is a better alternative matedal for urinary tissue engineering. OBJECTIVE： To construct the compound of hair folliole stem cells with heterologous bladder acellular scaffold, and to observe the growth of hair follicle stem cells on the scaffold; METHODS： Bladder acellular matrix from New Zealand rabbits were prepared and detected using scanning electron microscopy and Masson staining. Passage 3 hair follicle stem cells were statically inoculated into the surface of bladder acellular matrix using secondary sedimentation method. Under inverted microscope, cell growth was observed, and cell growth curves were drawn. Cell growth on the scaffold surface was observed through histological detection and scanning electron microscope observation. RESULTS AND CONCLUSION： Prepared bladder aceUular matrix was a white translucent film with fiber mesh structure, and no residual cells were seen. Masson staining results indicated that the bladder acellular matrix had collagen structure, and no obvious residual cells. After culture for 48 hours, hair follicle stem cells grew well around the bladder acellular matrix under inverted microscope; 1 week later, hair follicle stem cells extended and adhered to the scaffold surface. These findings indicate that hair follicle stem cells have a good biocompatibility with the bladder aceltular matrix through in vitro culture.