CAJ | 학술논문

目的：建立大鼠骨髓间充质干细胞（ BMSCs ）体外分离、培养、羟基荧光素二醋酸盐琥珀酰亚胺脂（CFSE）荧光标记的实验方法。方法差速贴壁离心法分离纯化大鼠BMSCs，并进行表面标志物流式检测鉴定和成脂、成骨分化鉴定。 CFSE标记BMSCs ，荧光显微镜下观察BMSCs 形态和标记率，台盼蓝染色检测CFSE对BM-SCs存活率的影响，以筛选CFSE标记BMSCs的最佳条件。结果第5代BMSCs的特异性表面标志物： CD29阳性，阳性率为90．10％；CD90阳性，阳性率为96．61％；CD45阴性，仅0．40％表达。且BMSCs具有成脂和成骨分化能力。10μmol/L CFSE作用15 min时，BMSCs的CFSE标记率几乎达到100％，且对BMSCs 的活性没有影响（P＞0．05）。结论利用差速贴壁法可获得纯度高、未分化性能强的BMSCs，BMSCs标记CFSE的最佳条件为10μmol/L CFSE作用15 min。
목적：건립대서골수간충질간세포（ BMSCs ）체외분리、배양、간기형광소이작산염호박선아알지（CFSE）형광표기적실험방법。방법차속첩벽리심법분리순화대서BMSCs，병진행표면표지물류식검측감정화성지、성골분화감정。 CFSE표기BMSCs ，형광현미경하관찰BMSCs 형태화표기솔，태반람염색검측CFSE대BM-SCs존활솔적영향，이사선CFSE표기BMSCs적최가조건。결과제5대BMSCs적특이성표면표지물： CD29양성，양성솔위90．10％；CD90양성，양성솔위96．61％；CD45음성，부0．40％표체。차BMSCs구유성지화성골분화능력。10μmol/L CFSE작용15 min시，BMSCs적CFSE표기솔궤호체도100％，차대BMSCs 적활성몰유영향（P＞0．05）。결론이용차속첩벽법가획득순도고、미분화성능강적BMSCs，BMSCs표기CFSE적최가조건위10μmol/L CFSE작용15 min。
Objective To investigate the method of isolation , cultivation and CFSE label of rat bone marrow mesenchymal stem cells (BMSCs).Methods BMSCs were isolated and expansion by adherent method .The specific surface markers of fifth generation BMSCs :CD29, positive rate was 90.10%.CD90, positive rate was 96.61%.CD45, negative rate was 0.40%, via flow cytometry, and differentiation.BMSCs labeled with CFSE were observed under fluorescence microscope and the labeling efficiencies were assessed .The effect of CFSE on the survival of BMSCS were tested by trypanblue .Re-sults FCM detected that CD 29 and CD90 were positively expressed , but CD34 was negatively expressed in BMSCs .Follow-ing induction, oil red O staining and alizarin red staining produced a strong reaction in cells .Nearly 100 % BMSCs were CFSE-labeled when BMSCs reacted with 10 μmol/L CFSE for 15 min.At the same time, the highest viability of BMSCs were observed(P>0.05).Conclusion BMSCs were obtained by adherent method .And staining with CFSE at 10μmol/L for 15 min was the optimal condition for rat BMSCs labeling in vitro .