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目的：研究大鼠骨髓间充干细胞（rBMSC）抵抗人血清介导的异种体液性杀伤的作用及其主要机制。方法：分离培养SD 大鼠BMSC，将第4 代的rBMSC 作为实验材料，以大鼠淋巴细胞（rLC）作为对照。采用流式细胞技术，检查两种细胞异种抗原alpha-Gal 的表达情况，体积分数20%正常人血清对两种细胞的杀伤作用、两种细胞分别与正常人血清中天然抗体IgM 和IgG的结合情况，以及与正常人血清作用后补体C3c、C4c、C5b-9 在两种细胞上的沉积情况。结果：成功分离和培养rBMSC。相对于rLC，rBMSC 对人血清介导的异种体液性杀伤具有明显的抵抗作用（P＜0.01）；rBMSC 上alpha-Gal的表达显著低于rLC（P＜0.05）；rBMSC 与人血清中IgG和IgM的结合量显著低于rLC（P＜0.01）；与正常人血清作用后，rLC 可见显著的C3c、C4c 和C5b-9 沉积，但rBMSC仅有较少C3c 和C4c 沉积，两种细胞间比较，差异均有统计学意义（P＜0.01）。结论：rBMSC 能明显抵抗人血清介导的异种体液免疫杀伤作用，其机制可能与rBMSC 低表达异种抗原琢-Gal及抑制了补体攻膜复合物形成有关。

英文摘要:

Objective:To investigate whether rat bone marrow mesenchymal stem cells (rBMSC) could resist human xenoantibody-dependant complement-mediated lysis and to explore its possible mechanisms.Methods:SD rat BMSCs were isolated. The rBMSCs at passage 4 were used for the following studies and rat lymphocytes were harvested as control cells. alpha-Gal expression was detected by flow cytometry. After incubation of rBMSCs with 20%normal human serum (NHS) or heat inactivated normal human serum (HINHS), flow cytometry was used to detect cytotoxicity, IgG or IgM binding, and C3c, C4c and C5b-9 deposition.Results:We successfully established the method to isolate and culture rBMSCs. As compared with rLCs, rBMSCs significantly resisted human natural antibody and complement-mediated lysis when incubated with 20% NHS. Mechanistically, rBMSCs expressed lower level of alpha-Gal, which was correlated with decreased binding of human xenoreactive IgG and IgM, and reduced deposition of complements C3c, C4c and C5b-9.Conclusion:These data demonstrated that the resistance of rBMSCs to human xenoantibody and complement-mediated lysis is associated with low expression of xenoaatigen alpha-Gal and inbibition of MAC (membrance attack complex) formation.

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