CAJ | 학술논문

目的探索骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)藻酸钙复合凝胶与经5-氮杂胞苷诱导的肌样细胞藻酸钙复合凝胶在压力性尿失禁(stress urinary incontinence,SUI)大鼠尿道周围的成肌效应.方法 SD大鼠BMSC体外分离、培养、鉴定；用5-氮杂胞苷诱导生成肌样细胞；2％藻酸钠与1％氯化钙溶液以5∶1体积比配制藻酸钙凝胶,分别与BMSC和肌样细胞复合用于微量注射.72只6周龄雌性SD大鼠建立SUI模型后分为BMSC凝胶组、肌样细胞凝胶组、单纯凝胶组和空白对照4组,每组再分为3小组,于膀胱颈尿道移行部黏膜下肌层注射相应的复合凝胶.4周、8周时取各组大鼠尿道横截面进行HE染色、荧光示踪照相以及结蛋白、α-横纹肌动蛋白(α-SMA)染色检查. 结果获得的BMSC第3代细胞表面细胞因子抗体CD29阳性率为89.4％、CD34为3.3％、CD45为2.5％、CD105为46.0％,荧光示踪照相见第3代培养细胞表达强绿色荧光.BMSC凝胶组和肌样细胞凝胶组4周、8周时,凝胶边缘血管长入并逐渐增多,荧光示踪BMSC聚集于新生血管周围,肌样细胞呈长梭形样生长,结蛋白、d-SMA明显阳性表达. 结论 BMSC、肌样细胞藻酸钙复合凝胶在大鼠SUI实验模型的微环境中具有向肌细胞分化的潜力.BMSC直接体内微环境诱导分化与5-氮杂胞苷体外诱导形成肌样细胞后植入体内微环境继续分化,短期结果无明显差异.
목적 탐색골수간충질간세포(bone marrow mesenchymal stem cell,BMSC)조산개복합응효여경5-담잡포감유도적기양세포조산개복합응효재압력성뇨실금(stress urinary incontinence,SUI)대서뇨도주위적성기효응.방법 SD대서BMSC체외분리、배양、감정；용5-담잡포감유도생성기양세포；2％조산납여1％록화개용액이5∶1체적비배제조산개응효,분별여BMSC화기양세포복합용우미량주사.72지6주령자성SD대서건립SUI모형후분위BMSC응효조、기양세포응효조、단순응효조화공백대조4조,매조재분위3소조,우방광경뇨도이행부점막하기층주사상응적복합응효.4주、8주시취각조대서뇨도횡절면진행HE염색、형광시종조상이급결단백、α-횡문기동단백(α-SMA)염색검사. 결과 획득적BMSC제3대세포표면세포인자항체CD29양성솔위89.4％、CD34위3.3％、CD45위2.5％、CD105위46.0％,형광시종조상견제3대배양세포표체강록색형광.BMSC응효조화기양세포응효조4주、8주시,응효변연혈관장입병축점증다,형광시종BMSC취집우신생혈관주위,기양세포정장사형양생장,결단백、d-SMA명현양성표체. 결론 BMSC、기양세포조산개복합응효재대서SUI실험모형적미배경중구유향기세포분화적잠력.BMSC직접체내미배경유도분화여5-담잡포감체외유도형성기양세포후식입체내미배경계속분화,단기결과무명현차이.
Objective To explore the effects of myoblast formation around the urethra of stress urinary incontinence (SUI) rats after treated with bone marrow mesenchymal stem cells(BMSCs) or musclelike cells/calcium alginate composite gel injection therapy. Methods Isolation,cultivation and identification of Sprague-Dawley rat bone marrow mesenchymal stem cell were performed.5-azacytidine was introduced to induce muscle-like cells.Calcium alginate gel was initially prepared by 2％ sodium alginate and 1％ calcium chloride solution at a volume ratio of 5∶ 1.Compounds of stem cells or muscle-like cells were mixed with gel,respectively,and were prepared for microinjection.SUI was produced in 72 6-week-old female Sprague-Dawley rats.The rats were then divided into 4 groups:Gel group,stem cell-gel group,muscle-like cell-gel group and mock control group.Each group was further divided into 3 groups.Submucosal injection of gel was performed at urethra and bladder neck.After preparation of cross sections of rat urinary tract at 4 weeks and 8 weeks after injection,HE staining,fluorescent tracing,staining of Desmin and α-skeletal muscle actin (α-SMA) were performed.OD values of positive rates were compared. Results At 4 weeks and 8 weeks after injection in stem cell-gel group and muscle-like cell-gel group,growth of blood vessels gradually increased at gel edge,BMSCs and muscle-like cells gathered around the new blood vessels observed by fl(u)orescence tracer,muscle-like cells grew into elongated spindle-like cells.Desmin and α-SMA staining were positive in these groups,and the OD values in the stem cell-gel group and muscle-like cell-gel group was significantly higher than that from the gel only group and control group,but no difference was found between stem cell-gel group and muscle-like cell-gel group. Conclusions Compound of BMSCs,muscle-like cells and calcium alginate composite gel has the potential to differentiate into muscle cells in the microenvironment of SUI rat model.In short term,the myoblast formation potential is the same whether the BMSCs was introduced into the micro-environment in vivo directly,or the BMSCs was implanted into microenvironment after the formation of the muscles cells induced by 5-azacytidine in vitro.