采用异种真皮去细胞基质复合脂肪间充质干细胞行尿道重建修复的实验研究  

Heterogenous acellular dermal matrix seeded with adipose derived stem cells for urethral reconstruction in a rabbit model

作　　者：黄炳伟[1] 李森[1] 喻希[1] 葛鹏[1] 林健[1] 

机构地区：[1]北京大学第一医院泌尿外科北京大学泌尿外科研究所,100034

出　　处：《中华泌尿外科杂志》2013年第10期787-792,共6页Chinese Journal of Urology

摘　　要：目的探讨异种真皮去细胞基质（acellular dermal matrix，ADM）种植脂肪间充质干细胞（adipose derived stemceils，ADSC）在兔的尿道重建修复中应用的效果。方法2012年3—11月选取成年雄性新西兰白兔12只，体质量2．5～4．0kg。体外构建长约1cm的尿道缺损模型，随机分为实验组和对照组，每组6只。无菌条件下手术获取实验组的脂肪组织，体外原代培养获取第3代ADSCs，通过细胞形态学和流式细胞术对其进行鉴定；将ADSC种植于人来源的ADM上制成组织工程化尿道。实验组用组织工程化尿道行尿道重建修复，对照组单纯使用ADM重建修复。于术后3、6个月时，每组分别取3只行尿道造影后处死并取修复处尿道行组织学检查，通过HE染色及免疫组化染色方法，使用抗兔细胞角蛋白AE1／AE3抗体检测尿路上皮细胞生长情况及抗兔肌间蛋白抗体检测平滑肌细胞生长情况，评估尿道缺损重建修复的效果。结果光镜下见分离培养的ADSC呈不规则长梭形，大小较为均匀，细胞间有交互联系，呈巢状生长。流式细胞术鉴定：分离培养的ADSC阳性表达CD166、CD105、CD90、CD44阴性表达CD45、CD13两组术后均可通过尿道排尿，尿路造影检查示无狭窄形成。术后3个月组织学检查示实验组修复材料上有平滑肌细胞长人，对照组则未见平滑肌细胞，两组修复材料上均未见上皮细胞生长。术后6个月组织学检查示两组修复材料上均有平滑肌细胞长入，但对照组平滑肌细胞较少；实验组修复材料上可见上皮细胞生长，形成尿路上皮细胞层；对照组未见上皮细胞生长。结论应用组织工程学方法，在异种的ADM上种植ADSCs制成组织工程化尿道进行尿道缺损修复，可促进缺损处尿道的再生，改善尿道修复的效果。Objective To assess the feasibility of heterogeneous acellular dermal matrix （ADM） seeded with adipose derived stem cells （ADSC） for urethroplasty in a rabbit model. Methods ADSC were isolated from a rabbit and expanded in vitro, then identified by flow cytometry. We seeded ADSC onto the ADM and made it into tissue-engineered urethra. 12 male rabbits were removed 1 cm urethra and divided into experiment group and control group. There were 6 rabbits in each group. Reconstructed urethra with tissueengineered urethra was used in experiment group, while unseeded ADM were used in control group. Urethrography was performed at 6 months after surgery. The animals were scarified at 3 and 6 months after surgery and the repaired urethra were harvested. H＆E staining and immunohistochemical staining were performed with cytokeratin AE1/AE3 and smooth muscle desmin makers. Results The morphology of isolated ADSC was with long spindle cross-links, and had multicentral growth. Flow cytometry showed that the ADSC expressed CD166, CD105, CO90 and CD44, but not expressed CD45 and CD13. The cells could growth well on the ADM and showed good biocompatibility with it. All animals could void normally, urethrography showed there was no significant stenosis. 3 months after surgery, the experiment group appeared regenerated smooth muscle hut not in the control group, both groups did not regenerate urothelium. At 6 months urothelium and smooth muscle cells could be observed in the experiment group, but only the smooth muscle was evident in the control gronp heterogeneous ADM effect of urethra. Conclusions By applying tissue engineering methods, we can seed the ADSC onto the and make it into tissue-engineered urethra, which can help improve the reconstructive

关 键 词：组织工程 尿道 脂肪间充质干细胞 真皮去细胞基质 

分 类 号：R699[医药卫生—泌尿科学]