人BMSCs分化过程中免疫能力的实验研究  

EXPERIMENTAL RESEARCH OF IMMUNE SUPPRESSION ABILITY OF HUMAN BONE MARROW MESENCHYMAL STEM CELLS DURING DIFFERENTIATION

作　　者：苏春燕[1] 苏鸿君[1] 王鹏[1] 冯佩[1] 吴燕峰[1] 

机构地区：[1]中山大学孙逸仙纪念医院生物治疗中心,广州510120

出　　处：《中国修复重建外科杂志》2015年第8期1003-1008,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81401850)~~

摘　　要：目的研究人BMSCs在成骨、成软骨及成脂分化过程中的免疫原性及其对外周血单个核细胞(peripheral blood mononuclear cell,PBMC)增殖的抑制能力。方法取健康志愿者骨髓分离培养BMSCs,分别进行成骨、成软骨和成脂诱导分化7、14、21 d。通过流式细胞术检测未分化及分化过程中BMSCs的人类白细胞抗原(human leukocyte antigen,HLA)Ⅰ类及Ⅱ类分子表达水平的改变。分离培养健康志愿者PBMC,按10∶1比例将PBMC与BMSCs共培养5 d,通过流式细胞术检测未分化及分化过程中BMSCs对PBMC增殖的抑制能力变化。结果未分化BMSCs表达HLA-Ⅰ类分子,基本不表达HLA-Ⅱ类分子。成骨、成软骨分化过程中各时间点HLA-Ⅰ、Ⅱ类分子表达无明显改变(P>0.05),且HLA-Ⅱ类分子表达水平均较低。成脂分化14、21 d HLA-Ⅰ类分子表达逐渐升高,与0、7 d比较差异有统计学意义(P<0.05);成脂分化7、14、21 d HLA-Ⅱ类分子表达也逐渐出现并升高,均显著高于0 d(P<0.05)。PBMC在无抗CD3、CD28微球刺激下无明显增殖,加入抗CD3、CD28微球后显著增殖,未分化BMSCs能显著抑制PBMC的增殖。成骨、成软骨分化过程中BMSCs抑制PBMC增殖能力无明显改变(P>0.05)。成脂分化7、14、21 d,BMSCs抑制PBMC增殖能力逐渐减弱,各时间点间差异均有统计学意义(P<0.05)。结论人BMSCs成骨、成软骨分化过程中仍具有低免疫原性和较强的免疫抑制能力,可作为同种异体组织工程修复和生物治疗的细胞;而成脂分化后免疫原性升高、免疫抑制能力降低,不适合作为同种异体组织工程修复和生物治疗的细胞。Objective To study the immunogenicity of human bone marrow mesenchymal stem cells（BMSCs） and the suppression ability to the proliferation of peripheral blood mononuclear cell（PBMC） during osteogenic, chondrogenic, and adipogenic differentiations. Methods BMSCs were isolated from bone marrow of healthy donors and were induced to osteogenic, chondrogenic, and adipogenic differentiations for 7, 14, and 21 days. The expressions of human leukocyte antigen（HLA） class I and class II were detected by flow cytometry. PBMC were isolated from peripheral blood of healthy donors and were co-cultured with BMSCs at a ratio of 10 ∶ 1 for 5 days. The suppression ability of undifferentiated and differentiated BMSCs to proliferation of PBMC were detected by flow cytometry. Results The HLA class I expression was observed but almost no expression of HLA class II was seen in undifferentiated BMSCs. There was no obviously change of the HLA class I and class II expressions during osteogenic and chondrogenic differentiations（P0.05）, and a low expression of HLA class II was kept. The HLA class I expression gradually increased at 14 and 21 days after adipogenic differentiation, showing significant differences when compared with the value at 0 and 7 days（P0.05）; the HLA class II expression also gradually increased at 7, 14, and 21 days after adipogenic differentiation, showing significant differences when compared with the value at 0 day（P0.05）. There was no proliferation of PBMC without the stimulation of CD3 and CD28 microspheres and significant proliferation was observed when CD3 and CD28 microspheres were added, and undifferentiated BMSCs could significantly inhibit the proliferation of PBMC. There was no obvious change of the ability of BMSCs to inhibit the proliferation of PBMC during osteogenic and chondrogenic differentiations（P0.05）; and the ability of BMSCs to inhibit the proliferation of PBMC was gradually weakened at 7, 14, and 21 days after adipogenic differentiation, showing significant difference

关 键 词：BMSCS 成骨分化 成软骨分化 成脂分化 免疫抑制能力 

分 类 号：R687[医药卫生—骨科学] R318.08[医药卫生—外科学]