不同造血发育阶段的人间充质干细胞研究
发布时间：2019-03-03 11:36
【摘要】： 目的:比较人骨髓(BM)、脐静脉(UV)、脐血(UCB)及体节期胚胎的背主动脉区(DA)来源的MSCs(mesenchymal stem cells, MSCs)的生物学特性,探讨MAPK及PI3K通路在BM-MSCs成骨分化中的作用。 方法:从人骨髓、脐静脉、脐血、DA中分离获得成纤维样细胞,对其进行细胞形态观察及细胞表型的检测。不同的诱导条件下,诱导其向脂肪、骨和软骨分化。进一步比较人骨髓、脐静脉、脐血来源的MSCs向不同谱系的分化能力及其对体外淋巴细胞增殖抑制能力。Western Blot检测MAPK/ERK及PI3K/AKT通路在BM-MSCs成骨过程中的变化,观察经PD98059及LY294002抑制该通路后对成骨的影响。 结果:从人骨髓、脐静脉、脐血及胚胎DA区分离得到的细胞具有以下特征:形态均一,为典型的纺锤状;表达CD105、CD73、间质细胞标志CD166、CD44、CD29及HLA-ABC;有成骨、成脂肪、成软骨分化的能力,符合MSCs的特征。比较骨髓、脐静脉、脐血三种来源的MSCs的结果显示,BM-MSCs成脂肪能力最强并且对异体淋巴细胞有较强的抑制能力;UV-MSCs成软骨能力较强,UCB-MSCs的成骨分化能力最强。在BM-MSCs向成骨分化过程中,早期即可检测到PI3K/Akt及MAPK/ERK通路被激活,加入特异性抑制剂后对成骨的抑制作用呈剂量依赖性。 结论:不同造血发育时期的组织,包括人体节期胚胎的AGM区、脐静脉、脐血和成体骨髓,均存在符合鉴定标准的MSCs。MAPK和PI3K通路参与BM-MSCs向成骨分化。
[Abstract]:Aim: to compare the biological characteristics of human bone marrow (BM), umbilical vein (UV), cord blood (UCB) and (DA)-derived MSCs (mesenchymal stem cells, MSCs) in dorsal aortic region of somatic phase embryo, and to explore the role of MAPK and PI3K pathway in BM-MSCs osteogenic differentiation. Methods: fibroblast-like cells were isolated from human bone marrow, umbilical vein, cord blood and DA. Under different induction conditions, it was induced to differentiate into fat, bone and cartilage. The ability of MSCs from human bone marrow, umbilical vein and cord blood to differentiate into different lineages and to inhibit the proliferation of lymphocytes in vitro were further compared. Western Blot was used to detect the changes of MAPK/ERK and PI3K/AKT pathways during the osteogenesis of BM-MSCs. To observe the effect of PD98059 and LY294002 on osteogenesis after inhibiting the pathway. Results: the cells isolated from human bone marrow, umbilical vein, cord blood and embryonic DA region showed the following characteristics: homogeneous morphology, typical spindle shape; expression of CD105,CD73, interstitial cell markers CD166,CD44,CD29 and HLA-ABC; Bone-forming, fat-forming, cartilage-forming ability to differentiate, in line with the characteristics of MSCs. The results of MSCs from bone marrow, umbilical vein and cord blood showed that BM-MSCs had the strongest fat-forming ability and strong inhibition ability to allogeneic lymphocytes, UV-MSCs had stronger cartilage-forming ability and UCB-MSCs had the strongest ability of osteogenic differentiation. The activation of PI3K/Akt and MAPK/ERK pathways can be detected early in the differentiation of BM-MSCs into osteogenesis, and the inhibition of osteogenesis by adding specific inhibitors is dose-dependent. Conclusion: in the tissues of different stages of hematopoietic development, including the AGM region of human ganglionic embryos, umbilical vein, cord blood and adult bone marrow, there are MSCs.MAPK and PI3K pathways that accord with the identification criteria and participate in the osteogenic differentiation of BM-MSCs.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329
本文编号：2433660
[Abstract]:Aim: to compare the biological characteristics of human bone marrow (BM), umbilical vein (UV), cord blood (UCB) and (DA)-derived MSCs (mesenchymal stem cells, MSCs) in dorsal aortic region of somatic phase embryo, and to explore the role of MAPK and PI3K pathway in BM-MSCs osteogenic differentiation. Methods: fibroblast-like cells were isolated from human bone marrow, umbilical vein, cord blood and DA. Under different induction conditions, it was induced to differentiate into fat, bone and cartilage. The ability of MSCs from human bone marrow, umbilical vein and cord blood to differentiate into different lineages and to inhibit the proliferation of lymphocytes in vitro were further compared. Western Blot was used to detect the changes of MAPK/ERK and PI3K/AKT pathways during the osteogenesis of BM-MSCs. To observe the effect of PD98059 and LY294002 on osteogenesis after inhibiting the pathway. Results: the cells isolated from human bone marrow, umbilical vein, cord blood and embryonic DA region showed the following characteristics: homogeneous morphology, typical spindle shape; expression of CD105,CD73, interstitial cell markers CD166,CD44,CD29 and HLA-ABC; Bone-forming, fat-forming, cartilage-forming ability to differentiate, in line with the characteristics of MSCs. The results of MSCs from bone marrow, umbilical vein and cord blood showed that BM-MSCs had the strongest fat-forming ability and strong inhibition ability to allogeneic lymphocytes, UV-MSCs had stronger cartilage-forming ability and UCB-MSCs had the strongest ability of osteogenic differentiation. The activation of PI3K/Akt and MAPK/ERK pathways can be detected early in the differentiation of BM-MSCs into osteogenesis, and the inhibition of osteogenesis by adding specific inhibitors is dose-dependent. Conclusion: in the tissues of different stages of hematopoietic development, including the AGM region of human ganglionic embryos, umbilical vein, cord blood and adult bone marrow, there are MSCs.MAPK and PI3K pathways that accord with the identification criteria and participate in the osteogenic differentiation of BM-MSCs.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329
【参考文献】
相关期刊论文 前1条
1 郭子宽,唐佩弦,刘晓丹,杨靖清,李秀森,陈小三,毛宁;人骨髓间充质干细胞支持体外造血(英文)[J];中国实验血液学杂志;2000年02期
，本文编号：2433660
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