CAJ | 학술논문

目的观察胎儿骨髓间充质干细胞(FBM-MSC)对成人外周血淋巴细胞的免疫调控作用及机制.方法取妊娠14～ 22周龄胎儿骨髓,分离、培养FBM-MSC,通过观察细胞形态、细胞表面分子标志及成骨、成脂分化潜能等进行鉴定;将FBM-MSC与用荧光染料CFSE标记的成人外周血单个核细胞,在PHA刺激条件下共培养5d后,用流式细胞术分析刺激活化后的淋巴细胞增殖情况;用ELISA法检测共培养上清中IFN-γ和TNF-α等炎性因子水平;用荧光实时定量RT-PCR法检测IDO、TSG-6和TGF-β等免疫抑制性因子表达水平.结果分离的FBM-MSC表面高表达CD29、CD44、CD49e、CD73、CD90、CD105;低表达CD31、CD34、CD45、HLA-DR,具有分化为脂肪和成骨细胞的能力;与单纯PHA刺激后的淋巴细胞相比较,与FBM-MSC共培养抑制淋巴细胞活化增殖,抑制率可高达96％; FBI-MSC共培养还可抑制淋巴细胞分泌炎性因子,对IFN-γ和TNF-α因子分泌的抑制率分别达90.9％和58.4％;与单独培养的FBM-MSC相比较,共培养后的FBM-MSC中重要的免疫调节因子如IDO、TSG-6和TGF-β的表达水平显著上调.结论胎儿骨髓来源的FBM-MSC对外周血淋巴细胞体外增殖具有明显抑制作用;由MSC分泌的抑制性细胞因子如IDO、TSG-6和TGF-β等在FBM-MSC介导的免疫抑制中可能起重要调节作用.
목적 관찰태인골수간충질간세포(FBM-MSC)대성인외주혈림파세포적면역조공작용급궤제.방법 취임신14～ 22주령태인골수,분리、배양FBM-MSC,통과관찰세포형태、세포표면분자표지급성골、성지분화잠능등진행감정;장FBM-MSC여용형광염료CFSE표기적성인외주혈단개핵세포,재PHA자격조건하공배양5d후,용류식세포술분석자격활화후적림파세포증식정황;용ELISA법검측공배양상청중IFN-γ화TNF-α등염성인자수평;용형광실시정량RT-PCR법검측IDO、TSG-6화TGF-β등면역억제성인자표체수평.결과 분리적FBM-MSC표면고표체CD29、CD44、CD49e、CD73、CD90、CD105;저표체CD31、CD34、CD45、HLA-DR,구유분화위지방화성골세포적능력;여단순PHA자격후적림파세포상비교,여FBM-MSC공배양억제림파세포활화증식,억제솔가고체96％; FBI-MSC공배양환가억제림파세포분비염성인자,대IFN-γ화TNF-α인자분비적억제솔분별체90.9％화58.4％;여단독배양적FBM-MSC상비교,공배양후적FBM-MSC중중요적면역조절인자여IDO、TSG-6화TGF-β적표체수평현저상조.결론 태인골수래원적FBM-MSC대외주혈림파세포체외증식구유명현억제작용;유MSC분비적억제성세포인자여IDO、TSG-6화TGF-β등재FBM-MSC개도적면역억제중가능기중요조절작용.
Objective To investigate the potential immunomodulatory properties of fetal bone marrow derived mesenchymal stem cells (FBM-MSCs).Methods Mononuclear cells from the bone marrow of second trimester (14-22 wks) fetus were isolated and cultured for the derivation of MSCs.The derived FBM-MSC cells were characterized via morphology,immunophenotyping and the adipogenic and osteogenic differentiation assays.The immunomodulatory properties of FBM-MSC on lymphocytes were evaluated through the co-culture assay with PHA activated adult peripheral blood mononuclear cells (PBMCs).Results Derived FBM-MSCs were CD29+,CD44,CD49e+,CD73,CD90+,CD105+ and CD31,CD34-,CD45,HLA-DR-and can be differentiated into adipocytes and osteocytes.When co-cultured with PHA-activated PBMCs,FBM-MSCs inhibited the proliferation of lymphocytes up to 96％ and down-regulated the secretion of inflammatory cytokines such as IFN-γ and TNF-α up to 90.9％ and 58.4％ respectively.When compared with FBM-MSCs cultured alone,the expression of MSCs derived immunomodulatory cytokines,such as IDO,TSG-6 and TGF-β,was up-regulated significantly in the co-culture system.Conclusions MSC derived from fetal bone marrow demonstrated immunosuppressive effects on adult PBMCs in vitro.MSC-derived cytokines like IDO,TSG-6 and TGF-β may be critical for FBM-MSCs mediated immunosuppressive function.