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  • 石磊,周伟东,马骏杰,等.脂肪间充质干细胞双向诱导上皮/平滑肌细胞膜片修复兔尿道缺损[J].同济大学学报(医学版),2020,41(5):561-566.    [点击复制]
  • SHI Lei,ZHOU Wei-dong,MA Jun-jie,et al.Bidirectional induction of adipose-derived mesenchymal stem cells into epithelial/smooth muscle cell membranes for repairing urethral defects in rabbits[J].同济大学学报(医学版),2020,41(5):561-566.   [点击复制]
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脂肪间充质干细胞双向诱导上皮/平滑肌细胞膜片修复兔尿道缺损
石磊,周伟东,马骏杰,李超,吴登龙
0
(同济大学医学院,上海200092;同济大学附属同济医院泌尿外科,上海200065)
摘要:
目的探究利用脂肪间充质干细胞(adipose-derived mesenchymal stem cells, ADSCs)向上皮/平滑肌细胞诱导培养细胞膜片重建兔缺损尿道的可行性和效果。方法将18只2~3月龄雄性新西兰兔随机分成ADSCs诱导复合膜片仿生尿道组和单纯无细胞SIS对照组,每组各9只。原代提取ADSCs体外扩增向上皮细胞和平滑肌细胞诱导并在UpCell培养皿中培养成上皮细胞膜片和平滑肌细胞膜片,两种细胞膜片叠加包裹8F医用硅胶管制备长2cm复合仿生尿道置皮下脂肪培养3周后取出。建立新西兰兔长段(2cm)尿道缺损模型,分别将ADSCs诱导复合细胞仿生尿道、单纯无细胞SIS仿生材料置于兔尿道缺损部位行全段端端吻合术修复重构尿道,术后14d移去8F医用硅胶管,分别于术后1、6个月进行尿道造影、大体观察和组织学观察,进行尿道重构效果评估。结果1个月后,尿道造影显示无细胞SIS材料组存在不同程度的尿道管腔狭窄,有明显的瘢痕增生和挛缩,而ADSCs诱导复合细胞仿生尿道组可保持通畅,黏膜平整,与正常尿道黏膜界限不清。ADSCs诱导复合细胞仿生尿道7只正常排尿畅通,2只出现尿瘘,无细胞SIS材料组4只正常排尿,3只尿道狭窄,2只出现尿瘘。组织学观察发现,6个月后ADSCs诱导复合膜片仿生尿道修复段已形成稳定的上皮细胞层多层组织结构,细胞层下可见明显的毛细血管样结构形成。结论ADSCs诱导双复合膜片仿生尿道可修复兔长段(2cm)尿道缺损的重构。
关键词:  脂肪间充质干细胞  诱导分化  细胞膜片  尿道重建
DOI:10.16118/j.1008-0392.2020.05.004
投稿时间:2020-03-07
基金项目:上海市科学技术委员会科研计划项目(18XD1403500)
Bidirectional induction of adipose-derived mesenchymal stem cells into epithelial/smooth muscle cell membranes for repairing urethral defects in rabbits
SHI Lei,ZHOU Wei-dong,MA Jun-jie,LI Chao,WU Deng-long
(Dept. of Ophthalmology, Yangpu Hospital, Tongji University School of Medicine, Shanghai 200090, China;Tongji University School of Medicine, Shanghai 200092, China; Dept. of Urology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China)
Abstract:
ObjectiveTo investigate the feasibility of induction of adipose-derived mesenchymal stem cells(ADSCs) into epithelial/smooth muscle cells for repairing urethral defects in rabbits. MethodsEighteen male New Zealand rabbits of 2-3 months were randomly divided into two groups with 9 rabbits in each group. In study group the urethral defects were repaired with ADSCs-induced composite patch and in control group the cell-free SIS was used for repairing. Primary extracted ADSCs were expanded in vitro to induce epithelial cells and smooth muscle cells and cultured into epithelial cell membrane and smooth muscle cell membrane in UpCell dish. The two cell membranes were superimposed and wrapped with 8F medical silica gel tube to prepare a 2cm-long composite biomimetic urethra and cultured in subcutaneous fat for 3 weeks before removal. The full-length(2cm) urethral defect model was established in New Zealand rabbits. ADSCs-induced composite cell biomimetic urethra(study group) and simple acellular SIS biomimetic material(control group) were placed in the urethral defect site of rabbits for full-segment end-to-end anastomosis to repair and reconstruct the urethra. 8F medical silicone catheter was removed 14 days after surgery. Urethrography, gross observation and histological observation were performed to evaluate the effect of urethral reconstruction 1 month and 6 months after surgery, respectively. ResultsOne month later, urethrography showed different degrees of urethral stricture, obvious scar hyperplasia and contracture in the control group; while in the study group, the urethra remained unobstructed, the mucosa was flat, and the boundary with normal urethral mucosa was unclear. In study group the normal voiding patency was found in 7 rabbits, urinary fistula in 2 rabbits; while in control group there were normal voiding in 4 rabbits, urethral stricture in 3, and urinary fistula in 2, respectively. Histological observation found that after 6 months, ADSCs induced biomimetic urethral repair segment with composite membrane had formed stable multi-layered tissue structure of epithelial cell layer, and obvious capillary-like structure formation was present under the cell layer. ConclusionThe biomimetic urethra induced by ADSCs with double-composite membranes can repair the defects and reconstruct the urethra with the whole length in rabbits.
Key words:  adipose-derived mesenchymal stem cells  induced differentiation  cell membrane  urethral reconstruction

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