免疫缺陷小鼠体内人源化骨髓微环境的重建  

Reconstruction of Humanized Bone Marrow Niche in Immunodeficiency Mouse

作　　者：陈欢 田征 李彬 邢海燕 李欢 唐克晶 王敏[1] 饶青 

机构地区：[1]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,天津300020

出　　处：《中国实验血液学杂志》2016年第6期1856-1862,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(81370599)

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。Objective：To reconstruct a human bone marrow niche in immunodeficiency mouse（NOD/SCID） so as to provide a model for observing the effect of abnormal BM niche on the occurence and development of leukemia.Methods：Human platelet lysate（HPL） was obtained by repeated freezing and thawing of concentrated platelet.Bone marrow-derived mesenchymal stem cells were cultured in α-minimal essential medium（α-MEM） containing 10%HPL or 10%FBS.The morphology,cell phenotype,multilineage differentiation potential in vitro and proliferation capacity between the mesenchymal stem cells cultured with HPL or FBS were compared.The human bone marrow formation capacity of HPL-cultured MSC was observed.The MSC was seeded on β-TCP scaffolds for 12 h,then the MSC-coated scaffold were implanted in a subcutaneous pocket on the dorsum of NOD/SCID mice.After 8-12 week,the scaffolds were harvested from the mice,then fixed,paraffin-embedded and stained for HE.Results：Whether cultured in the presence of HPL or FBS,the MSC all displayed a spindle-shaped fibroblast-like morphology;the flow cytometry analysis revealed no obvious differences in cell immunophenotype in this 2 groups;they all have the ability to differentiate towards osteoblasts,adipocytes,and chondrocytes in vitro.However,the mesenchymal stem cells cultivated with HPL-contained medium showed stronger proliferation capacity and higher activity to differentiate towards osteoblasts.Mesenchymal stem cells cultivated with HPL still have in vivo bone-forming capacity.Conclusion：HPL cultured MSC have stronger proliferation capacity and potential of differentiate towards osteoblasts,HPL-cultured MSC also can reconstruct humanized bone marrow niche in murine host.

关 键 词：人源骨髓微环境 骨髓间充质干细胞 人血小板裂解液 增殖能力 成骨分化 体内成骨 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]