组织工程化膀胱平滑肌结构的体外构建

Construction Of Tissue Engineering Bladder Smooth Muscle Structure In Vitro

目的利用组织工程方法在体外初步构建膀胱平滑肌结构。方法置备膀胱脱细胞基质移植物(BAMG)并以HE和Masson染色进行支架材料评价。酶消化法获得膀胱平滑肌原代细胞,分别以免疫荧光和RT-PCR进行细胞鉴定。经过体外培养和扩增后,将P3代膀胱平滑肌细胞接种在BAMG上。结果支架上的膀胱平滑肌细胞经过体外培养4周后形成多层结构,同时免疫组化检测α-SmoothMuscleActin表达阳性。结论运用组织工程方法能够在体外进行膀胱平滑肌结构的初步构建,为进一步膀胱组织工程修复奠定技术基础。

Objective To construct bladder smooth muscle structure in vitro by tissue engineering approach. Methods Fresh dog bladder was processed to fabricate bladder acellular matrix graft (BAMG) as a scaffold, which were evaluated by H&E and Masson's trichrome staining. Rabbit primary bladder smooth muscle cells(SMC) were harvested by collagen enzyme and confirmed with immunofluorescence and RT-PCR. After series culture and expansion in vitro, SMCs were reseeded in BAMG to form construction. Results SMCs were formed multilayer structure after 4 weeks culture in scaffold with α-smooth muscle actin positive staining by immunohistology. Conclusion Bladder smooth muscle structure can be constructed by tissue engineering approach. It will provide a technological basement for further bladder regeneration.