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目的探讨源于成人近端股骨的间充质干细胞(MSC)是否具有免疫抑制功能,为临床移植物抗宿主病的防治寻找理想的细胞储备库。方法从全髋关节置换术患者的近端股骨中分离并培养MSC,将不同数量的细胞直接或以Transwell法加入混合淋巴细胞反应(MLR)中,检测MSC对外周血淋巴细胞(PBL)增殖水平的影响。以ELISA法及抗体中和法探讨细胞因子TGFβ1在MSC免疫调节中的作用。结果3×103、1×104、3×104、1×105个源于近端股骨的MSC对PBL增殖的抑制率分别为62%±18%、38%±4%、31%±16%、28%±12%,与采自髂骨骨髓穿刺物MSC的抑制作用相比差异无统计学意义(均P>0.05),后者对PBL的增殖抑制率为58%±12%、37%±8%、28%±6%、27%±6%;以0.2μm的微孔膜对MSC与MLR进行分隔,3×104及1×104个MSC对PBL的增殖抑制率为36%±8%和53%±12%;在ELISA实验中,MSC单独培养,MSC与MLR直接接触,或经Transwell共培养的这3种上清中转化生长因子(TGF)β1的表达水平差异无统计学意义,加入浓度递增的TGFβ1抗体不能逆转MSC对PBL增殖的抑制作用。结论源于近端股骨的MSC具有一定的免疫抑制功能,除TGFβ1外其他可溶性因子可能介导这一作用。
Objective To investigate whether immunosuppressive function of mesenchymal stem cells (MSCs) derived from the proximal femur of adult rats is an ideal cell bank for the prevention and treatment of clinical graft-versus-host disease. Methods MSCs were isolated and cultured from the proximal femur of patients undergoing total hip arthroplasty. Different numbers of cells were added into mixed lymphocyte reaction (MLR) directly or by Transwell method to detect the proliferation of peripheral blood lymphocytes (PBL) Impact. To investigate the role of cytokine TGFβ1 in immune regulation of MSC by ELISA and antibody neutralization. Results The inhibitory rates of 3 × 103, 1 × 104, 3 × 104 and 1 × 105 MSC derived from the proximal femur on PBL proliferation were 62% ± 18%, 38% ± 4%, 31% ± 16% 28% ± 12%, respectively. There was no significant difference between the two groups (P> 0.05). The inhibition rates of the latter on PBL were 58% ± 12% and 37% ± 8%, 28% ± 6% and 27% ± 6%, respectively. MSC and MLR were separated by 0.2μm microporous membrane. The inhibition rate of 3 × 104 and 1 × 104 MSCs on PBL was 36% ± 8% And 53% ± 12% respectively. There was no significant difference in the expression of transforming growth factor (TGF) β1 among the three supernatants cultured in MSC alone or in MSC-MLR or co-cultured with Transwell in ELISA. Addition of TGFβ1 antibody in increasing concentrations did not reverse the inhibitory effect of MSC on PBL proliferation. Conclusion MSCs derived from proximal femur have some immunosuppressive functions. Other soluble factors other than TGFβ1 may mediate this effect.