CAJ | 학술논문

　　目的：研究不同剂量的3H-TdR对骨髓间充质干细胞生长代谢的影响，以确定安全的3H标记剂量。方法：在骨髓间充质干细胞培养瓶加入不同浓度的3H-TdR，描绘各细胞生长曲线，用Hochest染色观察细胞凋亡情况及用流式细胞仪检测细胞周期和骨髓间充质干细胞特有细胞表面标志物CD29、CD166、CD44、CD45的变化，观察对干细胞SOD、MDA的影响。结果：研究发现以4μCi/2ml浓度的3H-TdR标记干细胞时干细胞生长受到抑制，3μCi/2ml以上的浓度细胞均有不同程度的凋亡，流式细胞仪发现高浓度的核素能使干细胞被阻滞在S期及G2-M期，而且其细胞内的SOD下降及MDA升高，上述变化都有随核素剂量变化而变化的趋势；而用1μCi/2ml和正常组差异无统计学意义（P>0.05），标记过的骨髓间充质干细胞连续传代培养其增殖能力不受影响。结论：较高浓度的3H-TdR对干细胞的培养、示踪有影响，而1μCi/2ml的3H-TdR进行示踪研究是安全的。
　　목적：연구불동제량적3H-TdR대골수간충질간세포생장대사적영향，이학정안전적3H표기제량。방법：재골수간충질간세포배양병가입불동농도적3H-TdR，묘회각세포생장곡선，용Hochest염색관찰세포조망정황급용류식세포의검측세포주기화골수간충질간세포특유세포표면표지물CD29、CD166、CD44、CD45적변화，관찰대간세포SOD、MDA적영향。결과：연구발현이4μCi/2ml농도적3H-TdR표기간세포시간세포생장수도억제，3μCi/2ml이상적농도세포균유불동정도적조망，류식세포의발현고농도적핵소능사간세포피조체재S기급G2-M기，이차기세포내적SOD하강급MDA승고，상술변화도유수핵소제량변화이변화적추세；이용1μCi/2ml화정상조차이무통계학의의（P>0.05），표기과적골수간충질간세포련속전대배양기증식능력불수영향。결론：교고농도적3H-TdR대간세포적배양、시종유영향，이1μCi/2ml적3H-TdR진행시종연구시안전적。
Objective:To study the cellular toxicity of 3H-TdR in labeling mesenchymal stem cells (MSCs). Methods: Different dosage of 3H-TdR was added to the culture bottles. The growth curves of BMSC in different concentrations were made, the apoptosis of cell was observed by Hochest stain and the cell cycle was observed with flow cytometer. The cell surface marks of MSCs( CD29,CD166,CD44,CD45) were detected by flow cytometer and the influence on SOD or MDA level was observed. Results: The researches showed that MSCs growth was suppressed at 4μCi/2ml concentration of 3H-TdR. High concentration of 3H-TdR caused MSCs to be arrested at the S and G2-M stage; MSCs showed obvious apoptosis when the concentration of 3H-TdR exceeded 3μCi/2ml;SOD decreased,MDA increased and their changes were in proportion to the alteration of concentration of 3H-TdR. The proliferation ability of successive progeniture was not affected by 3H-TdR when its concentration didn't exceed 1μCi/2ml. Conclusion: Higher concentration of 3H-TdR can arrest cell cycle of stem cell,affect its growth and tracer function,but concentration lower than 1μCi/2ml has no effect on biological characteristic of stem cell.Therefore it is safe for tracing study.