CAJ | 학술논문

目的：探讨体外采用全骨髓贴壁法分离培养人骨髓间充质干细胞（human bone mesenchymal stem cells，hBMSCs）的可行性，为研究hBMSCs提供实验细胞来源。方法从因创伤导致骨折需行骨折复位及髂骨植骨术的志愿者中，获取骨髓，采用全骨髓贴壁法培养和分离hBMSCs，在倒置相差显微镜下观察细胞生长形态，取P3代细胞行表型鉴定；取P1、P2、P3代细胞，分别用胰蛋白酶消化，以培养时间为横轴，细胞数量为纵轴，绘制hBMSCs生长曲线图。结果经全骨髓贴壁法培养分离获得了纯度较高的hBMSCs，细胞呈均一的长梭形、短梭形及多角形，漩涡状或辐射状结构，绘制的生长曲线呈S形。P3代hBMSCs经流式细胞鉴定：CD29、CD44、CD90、CD105分别为99.6%、100%、99.4%、92.5%，呈阳性表达；而表达CD34、CD45分别为0.5%、0.6%，呈阴性表达。结论体外应用全骨髓贴壁培养法可成功分离hBMSCs，方法操作简单易行、经济，且能最大限度的保持细胞活力。
목적：탐토체외채용전골수첩벽법분리배양인골수간충질간세포（human bone mesenchymal stem cells，hBMSCs）적가행성，위연구hBMSCs제공실험세포래원。방법종인창상도치골절수행골절복위급가골식골술적지원자중，획취골수，채용전골수첩벽법배양화분리hBMSCs，재도치상차현미경하관찰세포생장형태，취P3대세포행표형감정；취P1、P2、P3대세포，분별용이단백매소화，이배양시간위횡축，세포수량위종축，회제hBMSCs생장곡선도。결과경전골수첩벽법배양분리획득료순도교고적hBMSCs，세포정균일적장사형、단사형급다각형，선와상혹복사상결구，회제적생장곡선정S형。P3대hBMSCs경류식세포감정：CD29、CD44、CD90、CD105분별위99.6%、100%、99.4%、92.5%，정양성표체；이표체CD34、CD45분별위0.5%、0.6%，정음성표체。결론체외응용전골수첩벽배양법가성공분리hBMSCs，방법조작간단역행、경제，차능최대한도적보지세포활력。
Objective To explore the feasibility of whole bone marrow adherent culture of human bone marrow mesenchymal stem cells (hBMSCs) in vitro. Methods The bone marrow was collected from the volunteers with traumatic fracture being reduced or from the volunteers receiving ilium graft;whole bone marrow adherent method was applied in culturing and isolating hBMSCs;inverted mi-croscope was used in observing the cell morphology and the cells of the 3rd generation was taken for cell phenotype identification;the cells of the first, the second and the third generation were taken and digested by trypsin respectively and hBMSCs growth curve was drawn with the culture time as the horizontal axis and the cell number as the longitudinal axis. Results hBMSCs cultured and isolated by whole bone marrow adherent method were of higher purity;the cells had long and short fusiform or in polygon, or in whorls or radial structure, the growth curve showed a typical"S"type;hBMSCs of the third generation were identified by flow cytometry:CD29, CD44, CD90 and CD105 were positive with the percentage of 99.6%, 100%, 99.4%and 92.5%respectively while CD 34 and CD45 were nega-tive with the percentage of 0.5% and 0.6% respectively. Conclusions Whole bone marrow adherent culture of hBMSCs in vitro is of easy operation and lower cost, more over, it can maintain maximum cell viability.