以兔毛囊干细胞与尿道黏膜干细胞为种子细胞构建组织工程尿道的比较  

Rabbit hair follicle stem cells and urethral mucosa stem cells used as seed cells for urethra tissue engineering: a comparison study

作　　者：彭文标[1] 刘春晓[1] 谢群[2] 朱臂琳 吕腾荣 

机构地区：[1]南方医科大学珠江医院泌尿外科,广州510515 [2]中山市博爱医院泌尿外科,中山528403 [3]中山市古镇医院泌尿外科,中山528400

出　　处：《第二军医大学学报》2013年第4期388-392,共5页Academic Journal of Second Military Medical University

基　　金：广东省自然科学基金(S2011010004029)~~

摘　　要：目的比较兔毛囊干细胞与尿道黏膜干细胞的表型及生物学特性,探索毛囊干细胞作为组织工程尿道种子细胞的可行性。方法体外分离、培养兔毛囊干细胞和尿道黏膜干细胞,并使用流式细胞仪分选富集直径<10μm、integrin-α6+/CD71-的干细胞。计算两种干细胞的最大扩增倍数和克隆形成能力,使用流式细胞仪检测两种干细胞K19、p63、β1-in-tegrin的阳性表达率。将富集毛囊干细胞和尿道黏膜干细胞接种在尿道支架上,构建组织工程尿道,应用组织学、荧光染色法观察体外构建的组织工程尿道的情况。结果兔毛囊干细胞与尿道黏膜干细胞的最大扩增倍数分别为(6.1±0.8)×104倍、(7.1±1.1)×103倍,克隆形成率分别为(10.1±1.3)%、(4.3±1.1)%。K19、p63、β1-integrin在毛囊干细胞中阳性表达率分别为(90.53±6.77)%、(93.31±5.57)%、(91.17±6.98)%,在尿道黏膜干细胞中阳性表达率分别为(88.50±4.95)%、(91.63±5.86)%、(93.35±6.28)%。毛囊干细胞与尿道黏膜干细胞在支架上都能形成复层上皮样结构,AE1/AE3染色阳性。结论兔毛囊干细胞与尿道黏膜干细胞都可以作为种子细胞构建组织工程尿道;在种子细胞获取方面,毛囊干细胞优于尿道黏膜干细胞。Objective To compare the phenotypes and biological characteristics between the rabbit hair follicle stem cells and urethral mucosa stem cells, and to explore the feasibility of using hair follicle stem cells as seed cells for tissue engineering urethra. Methods Rabbit hair follicle stem cells and urethral mucosa stem cells were isolated and cultured in vitro, and the cells with diameter〈10 μm and integrin-a6＋/CD71- were sorted using flow cytometry. The maximum amplification factor and clone forming ability of the two kinds of cells were calculated. The positive rates of K19, p63 and β1-integrin in the two kinds of cells were detected by flow cytometry. The enriched stem cells were implanted in the urethral stent to construct tissue engineering urethra. Histology and fluorescent staining were used to observe the tissue-engineered urethra. Results The maximum amplification multiples of rabbit hair follicle stem cells and urethral mucosa stem cells were （6.1±0.8） × 10^4 and （7.1±1.1）×10^3, respectively; and the clone formation rates were （10. 1±1. 3）% and （4. 3±1. 1）%, respectively. The positive rates of K19, p63, and β1-integrin in hair follicle stem ceils were （90.53±6.77） %, （93.31±5.57） %, and （91.17±6.98） %, respectively; and those in urethral mucosa stem cells were （88. 50 ±4. 95） %, （91. 63±5. 86） %, and （93. 35 ±6.28） %, respectively. Both hair follicle stem cells and urethral mucosa stem cells formed complex layer epithelioid structure on the stents, with positive staining for AE1/AE3. Conclusion Rabbit hair follicle stem cells and urethral mucosa stem cells can both serve as seed cells for constructing tissue engineering urethra. As for the availability of seed cells, hair follicle stem cells are more superior to the urethral mucosa stem cells.

关 键 词：毛囊干细胞 尿道黏膜干细胞 组织工程 尿道 

分 类 号：R695[医药卫生—泌尿科学]