Department of Urology, Zhongnan Hospital, Wuhan University, Wuhan, PR China.
J Surg Res. 2012 Nov;178(1):55-62. doi: 10.1016/j.jss.2012.01.047. Epub 2012 Mar 28.
The purpose of the present study was to assess the differentiation potential of adipose-derived stem cells (ASCs) into smooth muscle cells (SMCs) and their potential for promoting regeneration of smooth muscle for ureteral tissue engineering.
ASCs were isolated, proliferated, and identified in vitro. SMC differentiation was induced using SMC induction medium. Gene expression was evaluated by quantitative polymerase chain reaction, immunofluorescence, and Western blotting. Vessel extracellular matrix was obtained by a decellularization process. The induced cells were seeded onto vessel extracellular matrix for ureter reconstitution. Grafts were obtained for evolutionary histologic studies. Renal function and ureteral patency was evaluated by intravenous urography at 16 wk.
Flow cytometry demonstrated that the ASCs expressed CD90, but did not express CD45 or CD34. After 6 wk of induction, upregulation of α-smooth muscle actin expression was determined by quantitative polymerase chain reaction, and smooth muscle myosin heavy chain expression was confirmed by immunofluorescence and Western blotting in the induced cells. Vessel extracellular matrix exhibited a nontoxic and bioactive effect on the induced cells. Histologically, stratified urothelium and organized muscle bundles were observed in the grafts at 16 wk. Intravenous urography demonstrated no ureteral stricture or hydroureteronephrosis.
These results have demonstrated that ASCs can be differentiated into SMCs and this potential promoted smooth muscle regeneration for ureteral tissue engineering.
本研究旨在评估脂肪来源干细胞（ASCs）向平滑肌细胞（SMCs）分化的潜能及其在促进输尿管组织工程平滑肌再生中的应用。
体外分离、增殖和鉴定 ASCs。使用 SMC 诱导培养基诱导 SMC 分化。通过定量聚合酶链反应、免疫荧光和 Western blot 评估基因表达。通过脱细胞过程获得血管细胞外基质。将诱导的细胞接种到血管细胞外基质上，用于输尿管重建。获取移植物进行进化组织学研究。16 周时通过静脉尿路造影评估肾功能和输尿管通畅性。
流式细胞术显示 ASCs 表达 CD90，但不表达 CD45 或 CD34。诱导 6 周后，通过定量聚合酶链反应确定α-平滑肌肌动蛋白表达上调，免疫荧光和 Western blot 证实诱导细胞中平滑肌肌球蛋白重链表达。血管细胞外基质对诱导细胞表现出无毒和生物活性作用。组织学上，在 16 周的移植物中观察到分层尿路上皮和有组织的肌肉束。静脉尿路造影显示无输尿管狭窄或肾盂积水。
这些结果表明，ASCs 可分化为 SMCs，这种潜能促进了输尿管组织工程的平滑肌再生。
