【摘要】： 目的:比較人骨髓(BM)、臍靜脈(UV)、臍血(UCB)及體節(jié)期胚胎的背主動脈區(qū)(DA)來源的MSCs(mesenchymal stem cells, MSCs)的生物學(xué)特性,探討MAPK及PI3K通路在BM-MSCs成骨分化中的作用。 方法:從人骨髓、臍靜脈、臍血、DA中分離獲得成纖維樣細(xì)胞,對其進行細(xì)胞形態(tài)觀察及細(xì)胞表型的檢測。不同的誘導(dǎo)條件下,誘導(dǎo)其向脂肪、骨和軟骨分化。進一步比較人骨髓、臍靜脈、臍血來源的MSCs向不同譜系的分化能力及其對體外淋巴細(xì)胞增殖抑制能力。Western Blot檢測MAPK/ERK及PI3K/AKT通路在BM-MSCs成骨過程中的變化,觀察經(jīng)PD98059及LY294002抑制該通路后對成骨的影響。 結(jié)果:從人骨髓、臍靜脈、臍血及胚胎DA區(qū)分離得到的細(xì)胞具有以下特征:形態(tài)均一,為典型的紡錘狀;表達(dá)CD105、CD73、間質(zhì)細(xì)胞標(biāo)志CD166、CD44、CD29及HLA-ABC;有成骨、成脂肪、成軟骨分化的能力,符合MSCs的特征。比較骨髓、臍靜脈、臍血三種來源的MSCs的結(jié)果顯示,BM-MSCs成脂肪能力最強并且對異體淋巴細(xì)胞有較強的抑制能力;UV-MSCs成軟骨能力較強,UCB-MSCs的成骨分化能力最強。在BM-MSCs向成骨分化過程中,早期即可檢測到PI3K/Akt及MAPK/ERK通路被激活,加入特異性抑制劑后對成骨的抑制作用呈劑量依賴性。 結(jié)論:不同造血發(fā)育時期的組織,包括人體節(jié)期胚胎的AGM區(qū)、臍靜脈、臍血和成體骨髓,均存在符合鑒定標(biāo)準(zhǔn)的MSCs。MAPK和PI3K通路參與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.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R329

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相關(guān)期刊論文 前1條

1 郭子寬,唐佩弦,劉曉丹,楊靖清,李秀森,陳小三,毛寧;人骨髓間充質(zhì)干細(xì)胞支持體外造血(英文)[J];中國實驗血液學(xué)雜志;2000年02期

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