CAJ | 학술논문

背景:hoechst33342可以用于对骨髓间充质干细胞的标记,但是其标记的最佳浓度的研究目前尚未见报道.目的:观察不同浓度的Hoechst33342对大骨髓间充质干细胞的增殖、标记率的影响,以及其荧光持续时间.方法:贴壁筛选提取大鼠骨髓间充质干细胞,培养至第3代,流式细胞仪检测其表面抗原,成骨、成脂诱导后染色,用不同浓度Hoechst33342标记大鼠骨髓间充质干细胞后,检测标记后的大鼠骨髓间充质干细胞增殖率、标记率,荧光显微镜下观察其持续时间.结果与结论:① hoechst33342可以用于骨髓间充质干细胞的标记,其最佳标记浓度为5 mg/L,标记持续时间较长,30 d未见猝灭.②其荧光激发后对细胞有损伤作用,标记浓度超过10 mg/L时,可致细胞死亡.③标记浓度在7.5 mg/L时,细胞增殖受到抑制;标记浓度在2.5 mg/L 时,标记时间较短,7 d 时荧光减弱,14 d 荧光消失.结果表明, hoechst33342可用于骨髓间充质干细胞的标记,其对骨髓间充质干细胞标记的最佳浓度为5 mg/L;细胞表型鉴定CD45阴性,CD44,CD90均呈阳性表达.
배경:hoechst33342가이용우대골수간충질간세포적표기,단시기표기적최가농도적연구목전상미견보도.목적:관찰불동농도적Hoechst33342대대골수간충질간세포적증식、표기솔적영향,이급기형광지속시간.방법:첩벽사선제취대서골수간충질간세포,배양지제3대,류식세포의검측기표면항원,성골、성지유도후염색,용불동농도Hoechst33342표기대서골수간충질간세포후,검측표기후적대서골수간충질간세포증식솔、표기솔,형광현미경하관찰기지속시간.결과여결론:① hoechst33342가이용우골수간충질간세포적표기,기최가표기농도위5 mg/L,표기지속시간교장,30 d미견졸멸.②기형광격발후대세포유손상작용,표기농도초과10 mg/L시,가치세포사망.③표기농도재7.5 mg/L시,세포증식수도억제;표기농도재2.5 mg/L 시,표기시간교단,7 d 시형광감약,14 d 형광소실.결과표명, hoechst33342가용우골수간충질간세포적표기,기대골수간충질간세포표기적최가농도위5 mg/L;세포표형감정CD45음성,CD44,CD90균정양성표체.
BACKGROUND:Hoechst33342 can be used for labeling bone marrow mesenchymal stem cel s, but there are few reports on the optimal concentration of marker. OBJECTIVE:To investigate the effect of different concentrations of Hoechst33342 on the proliferation and labeling rate of rat bone marrow mesenchymal stem cel s and to investigate the duration time of fluorescence. METHODS:Adherence screening method was used to extract rat bone marrow mesenchymal stem cel s, and the cel s were cultured to the third generation, then surface antigen was detected by flow cytometry, and stained after osteogenic and adipogenic induction. The proliferation rate and labeling rate of rat bone marrow mesenchymal stem cel s were detected after treatment with different concentrations of Hoechst33342, and then the duration time was detected through the use of fluorescence microscope. RESULTS AND CONCLUSION:Hoechst33342 can be used for labeling bone marrow mesenchymal stem cel s, and the optimal concentration of Hoechst33342 was 5 mg/L, the duration time was long and there was no quenching within 30 days. Hoechst33342 damaged the cel s after fluorescence excitation, and when the concentrations exceed10 mg/L, the marker could cause cel death. When the concentration was 7.5 mg/L, cel proliferation was inhibited;when the concentration was 2.5 mg/L, the duration for labeling was shorter, the fluorescence decreased at 7 days and disappeared at 14 days. Hoechst33342 can be used for labeling bone marrow mesenchymal stem cel s, and the optimal concentration of the Hoechst33342 is 5 mg/L;phenotype identification shows the negative expression of CD45 and positive expression of CD44 and CD90.