免疫缺陷小鼠体内人源化骨髓微环境的重建

发布时间：2018-11-11 12:57

【摘要】：目的:通过在免疫缺陷小鼠体内重建人源的骨髓微环境,为进一步研究人异常的骨髓微环境在人白血病发生发展中的作用提供模型。方法:通过反复冻融浓缩后血小板获得人血小板裂解液(HPL),以α-MEM作为基础培养液分别添加10%HPL和10%胎牛血清(FBS)培养骨髓来源的间充质干细胞。比较两种添加成分对MSC形态、免疫表型、多向分化能力以及增殖能力的影响;将HPL培养的MSC接种于β-TCP支架材料并移植到免疫缺陷小鼠背部皮下,8-12周后取出移植体做HE染色,观察MSC在体内形成骨以及骨髓结构的能力。结果:用HPL和FBS培养的M SC均呈现长梭形纤维样细胞结构,均具有多向分化能力;两种体系培养的M SC免疫表型无明显区别,但用人血小板裂解液培养的MSC具有更强的增殖能力和向成骨分化能力;用人血小板裂解液培养的MSC具有在体内形成骨组织样结构的能力。结论:HPL培养的MSC具有更强的增殖能力以及成骨分化潜能,它能在NOD/SCID鼠上重建人源化骨髓微环境。
[Abstract]:Aim: to provide a model for the further study of the role of human bone marrow microenvironment in the development of human leukemia by reconstructing the human bone marrow microenvironment in immunodeficient mice. Methods: human platelet lysate (HPL), supplemented with 10%HPL and 10% fetal bovine serum (FBS) supplemented with 伪-MEM was used to culture mesenchymal stem cells derived from bone marrow by repeatedly freezing and thawing concentrated platelets. The effects of two additive components on the morphology, immunophenotype, multidirectional differentiation and proliferation of MSC were compared. The MSC cultured by HPL was inoculated into 尾-TCP scaffold and transplanted into the subcutaneous of the back of immunodeficient mice. After 8-12 weeks, the grafts were removed for HE staining to observe the ability of MSC to form bone and bone marrow structure in vivo. Results: M SC cultured with HPL and FBS showed long fusiform fibrous cell structure and multidirectional differentiation ability. There was no significant difference in the immunophenotype of M SC between the two systems, but the MSC cultured in human platelet lysate had stronger proliferative and osteogenic ability. MSC cultured with human platelet lysate has the ability to form bone-like structure in vivo. Conclusion: MSC cultured with HPL has stronger proliferative and osteogenic potential, and it can reconstruct human bone marrow microenvironment in NOD/SCID mice.
【作者单位】：中国医学科学院北京协和医学院血液学研究所血液病医院实验血液学国家重点实验室;
【基金】：国家自然科学基金资助项目(81370599)
【分类号】：R593.3

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