CAJ | 학술논문

目的探讨脂肪干细胞与聚羟基乙酸(PGA)材料复合在体外诱导构建尿路肌性管腔的可行性.方法酶消化法获取脂肪干细胞,经体外培养和扩增后,流式细胞技术检测细胞表面抗原CD90、CD105、CD34,并接种于PGA上,形成细胞-材料复合物,体外培养4周后,采用由10 μmol／L5-氮杂胞苷+5％马血清+5％胎牛血清组成的成肌诱导培养液诱导4周,行大体观察及组织学检测,免疫组织化学法检测成肌细胞特异性抗原结蛋白(desmin)和肌球蛋白(myosin)表达.结果流式细胞技术检测CD90、CD105及CD34的阳性率分别为94.2％、90.7％和0.6％,所构建组织形成类似尿道的肌性管腔结构,管腔轻度塌陷,免疫组织化学染色显示:脂肪干细胞在诱导4周后,表达desmin和myosin;细胞材料复合物胶原成分较多.结论脂肪干细胞复合PGA后生长良好,可在体外完成成肌诱导,构建肌性管腔.
목적 탐토지방간세포여취간기을산(PGA)재료복합재체외유도구건뇨로기성관강적가행성.방법 매소화법획취지방간세포,경체외배양화확증후,류식세포기술검측세포표면항원CD90、CD105、CD34,병접충우PGA상,형성세포-재료복합물,체외배양4주후,채용유10 μmol／L5-담잡포감+5％마혈청+5％태우혈청조성적성기유도배양액유도4주,행대체관찰급조직학검측,면역조직화학법검측성기세포특이성항원결단백(desmin)화기구단백(myosin)표체.결과 류식세포기술검측CD90、CD105급CD34적양성솔분별위94.2％、90.7％화0.6％,소구건조직형성유사뇨도적기성관강결구,관강경도탑함,면역조직화학염색현시:지방간세포재유도4주후,표체desmin화myosin;세포재료복합물효원성분교다.결론 지방간세포복합PGA후생장량호,가재체외완성성기유도,구건기성관강.
Objective To investigate the feasibility of in vitro induction constructing the muscular conduit of urinary tract engineered from adipose-derived stem cells (ADSCs) seeded on polyglycolic acid (PGA).Methods ADSCs were harvested by collagen enzyme.After series culture and expansion in vitro,the phenotypic analyses of ADSCs including CD90,CD105 and CD34 were identified by flow cytometry,and cell-PGA complex was constructed by seeding ADSCs on PGA fibers.After in vitro culture for 4 weeks,the induced culture medium containing 10 μmol/L 5-azacytidine (5-aza),5％ horse serum and 5％ fetal bovine serum was used to induce cell-PGA complex for four weeks,and the engineered tissue were examined both grossly and histologically.The immunohistochemistry was utilized to detect the expression of myoblast specific antigens desmin and myosin.Results ADSCs were positive for CD90 (94.2％) and CD105 (90.7％),lacking in CD34 (0.6％).The engineered tissue was similar to the muscular conduit of urinary tract,and its lumen collapsed slightly.Immunohistochemistry staining showed that ADSCs,after induction for 4 weeks,expressed desmin and myosin.There was much collagen in cell-PGA complex.Conclusion ADSCs have good biocompatibility with PGA and can differentiate into myoblasts.The cell-PGA complex can construct the muscular conduit of urinary tract in vitro.