【摘要】：目的研究人BMSCs在成骨、成軟骨及成脂分化過程中的免疫原性及其對(duì)外周血單個(gè)核細(xì)胞(peripheral blood mononuclear cell,PBMC)增殖的抑制能力。方法取健康志愿者骨髓分離培養(yǎng)BMSCs,分別進(jìn)行成骨、成軟骨和成脂誘導(dǎo)分化7、14、21 d。通過流式細(xì)胞術(shù)檢測(cè)未分化及分化過程中BMSCs的人類白細(xì)胞抗原(human leukocyte antigen,HLA)Ⅰ類及Ⅱ類分子表達(dá)水平的改變。分離培養(yǎng)健康志愿者PBMC,按10∶1比例將PBMC與BMSCs共培養(yǎng)5 d,通過流式細(xì)胞術(shù)檢測(cè)未分化及分化過程中BMSCs對(duì)PBMC增殖的抑制能力變化。結(jié)果未分化BMSCs表達(dá)HLA-Ⅰ類分子,基本不表達(dá)HLA-Ⅱ類分子。成骨、成軟骨分化過程中各時(shí)間點(diǎn)HLA-Ⅰ、Ⅱ類分子表達(dá)無明顯改變(P0.05),且HLA-Ⅱ類分子表達(dá)水平均較低。成脂分化14、21 d HLA-Ⅰ類分子表達(dá)逐漸升高,與0、7 d比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);成脂分化7、14、21 d HLA-Ⅱ類分子表達(dá)也逐漸出現(xiàn)并升高,均顯著高于0 d(P0.05)。PBMC在無抗CD3、CD28微球刺激下無明顯增殖,加入抗CD3、CD28微球后顯著增殖,未分化BMSCs能顯著抑制PBMC的增殖。成骨、成軟骨分化過程中BMSCs抑制PBMC增殖能力無明顯改變(P0.05)。成脂分化7、14、21 d,BMSCs抑制PBMC增殖能力逐漸減弱,各時(shí)間點(diǎn)間差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論人BMSCs成骨、成軟骨分化過程中仍具有低免疫原性和較強(qiáng)的免疫抑制能力,可作為同種異體組織工程修復(fù)和生物治療的細(xì)胞;而成脂分化后免疫原性升高、免疫抑制能力降低,不適合作為同種異體組織工程修復(fù)和生物治療的細(xì)胞。
[Abstract]:Objective to study the immunogenicity of human BMSCs in osteogenesis, cartilage formation and adipogenic differentiation and its inhibitory effect on the proliferation of peripheral blood mononuclear cells (peripheral blood mononuclear cell,PBMC). Methods BMSCs, was isolated and cultured from bone marrow of healthy volunteers for osteogenesis, cartilage formation and adipogenic differentiation for 7, 14 and 21 days, respectively. The expression of human leukocyte antigen (human leukocyte antigen,HLA (human leukocyte antigen,HLA) class I and class II in undifferentiated and differentiated BMSCs was detected by flow cytometry. PBMC, was co-cultured with BMSCs for 5 days in the proportion of 10:1. Flow cytometry was used to detect the inhibitory effect of BMSCs on the proliferation of PBMC during undifferentiation and differentiation. Results HLA- class I molecules were expressed in undifferentiated BMSCs, but HLA- class II molecules were not expressed in undifferentiated HLA-. There was no significant change in the expression of HLA- class 鈪，

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