骨髓间充质干细胞对白血病细胞生长影响的研究

发布时间：2018-01-05 22:05

本文关键词：骨髓间充质干细胞对白血病细胞生长影响的研究　出处：《南京医科大学》2009年硕士论文　论文类型：学位论文

【摘要】：目的比较K562细胞与相同数量以及不同数量的人骨髓间充质干细胞(MSCs)粘附培养前后K562细胞增殖、细胞周期和凋亡的变化,以研究骨髓间充质干细胞对人白血病K562细胞生长的影响。方法通过建立正常人骨髓间充质干细胞的体外培养体系及其与K562细胞共培养的体系测定K562细胞的生长曲线;将不同数量的MSCs与K562细胞共培养测定K562细胞的增殖率曲线;流式细胞仪检测K562细胞周期以及凋亡的变化。结果与单独K562细胞培养相比,与相同数量MSCs共培养后,K562细胞生长受抑;与不同数量MSCs共培养后,其对K562细胞的抗增殖作用呈剂量依赖性;细胞周期分析发现K562细胞与MSCs共培养后,G0/G1期以及G2/M期的细胞增加, S期的细胞减少。共培养24 h、48 h、72 h后用流式细胞仪检测其凋亡率,发现K562细胞的凋亡率下降。结论正常人骨髓间充质干细胞能导致K562细胞生长抑制,阻止K562细胞周期的运行,凋亡率下降,且在一定范围内,随着间充质干细胞数量的增加其对K562细胞的抗增殖作用增强。目的观察骨髓间充质干细胞(BMMSC)对小鼠急性淋巴细胞白血病(ALL)模型中白血病细胞的作用; 方法将携带绿色荧光蛋白(eGFP)基因标记的P388细胞(小鼠急性淋巴细胞白血病细胞系)腹腔接种于DBA/2小鼠建立ALL小鼠动物模型,静脉输注体外培养的骨髓间充质干细胞。记录小鼠存活时间;监测小鼠血常规;流式细胞仪检测组织器官标本中的eGFP+细胞的百分比;观察各小鼠脏器发生的病理学变化以及检测输注MSC的小鼠体内Y染色体的存在。结果骨髓间充质干细胞能延长ALL小鼠的生存时间,但其血常规变化不大; MSCs能够减少各组织白血病细胞的浸润,并具有向各组织定植的能力。结论骨髓间充质干细胞能影响小鼠急性淋巴细胞白血病(ALL)模型中白血病细胞的生长。
[Abstract]:Purpose To compare the proliferation, cell cycle and apoptosis of K562 cells before and after adhesion culture with the same number and different numbers of human bone marrow mesenchymal stem cells (MSCs). To study the effect of bone marrow mesenchymal stem cells (BMSCs) on the growth of human leukemia K562 cells. Method The growth curve of normal human bone marrow mesenchymal stem cells (BMSCs) was determined by establishing the culture system in vitro and co-culture with K562 cells. The proliferation rate curve of K562 cells was measured by co-culture of different amounts of MSCs and K562 cells. The changes of K562 cell cycle and apoptosis were detected by flow cytometry. Results Compared with K562 cells alone, the growth of K562 cells was inhibited after co-culture with the same number of MSCs. The antiproliferative effect of MSCs on K562 cells was dose-dependent after co-culture with different amount of MSCs. Cell cycle analysis showed that K562 cells co-cultured with MSCs increased in G _ 0 / G _ 1 phase and G _ 2 / M phase, but decreased in S phase and co-cultured for 24 h or 48 h. After 72 hours, the apoptosis rate of K562 cells was detected by flow cytometry, and it was found that the apoptosis rate of K562 cells decreased. Conclusion Normal human bone marrow mesenchymal stem cells can induce K562 cell growth inhibition, prevent K562 cell cycle, apoptosis rate decreased, and within a certain range. As the number of mesenchymal stem cells increased, the anti-proliferation effect of mesenchymal stem cells on K562 cells was enhanced. Purpose To observe the effect of bone marrow mesenchymal stem cells (BMMSC) on leukemic cells in mice acute lymphoblastic leukemia (ALL) model. Method P388 cells (mouse acute lymphoblastic leukemia cell line) carrying green fluorescent protein (GFP) gene were inoculated intraperitoneally into DBA/2 mice to establish ALL mice model. Bone marrow mesenchymal stem cells (BMSCs) cultured in vitro were infused intravenously. Survival time of mice was recorded. The blood routine of mice was monitored. The percentage of eGFP cells in tissues and organs was detected by flow cytometry. The pathological changes of mouse organs and the existence of Y chromosome in mice injected with MSC were observed. Results Bone marrow mesenchymal stem cells (BMSCs) could prolong the survival time of ALL mice but had little change in blood routine. MSCs could reduce the infiltration of leukemic cells from various tissues and have the ability to colonize the tissues. Conclusion Bone marrow mesenchymal stem cells (BMSCs) can affect the growth of leukemic cells in acute lymphoblastic leukemia (ALL) model.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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