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目的:研究犬未分化骨髓基质干细胞(bone marrow stromal cells,BMSCs)体外抑制淋巴细胞增殖的作用。方法:从犬骨髓组织中分离获取骨髓基质干细胞,体外培养至第二代,1×105细胞/孔BMSCs刺激异体淋巴细胞,观察淋巴细胞增殖情况。以植物血凝素(PHA,2ug/mL)刺激淋巴细胞后,分别以1×102～5细胞/孔数量的异体BMSCs加入反应体系中,观察淋巴细胞增殖抑制情况。分别以1×102～5细胞/孔数量的“第三者”BMSCs加入双向混合淋巴细胞反应体系中,观察淋巴细胞增殖抑制情况。通透性隔离腔培养的BMSCs加入双向混合淋巴细胞反应体系中,观察淋巴细胞增殖抑制情况。结果:BMSCs刺激同种异体淋巴细胞体外增殖作用不明显;BM-SCs可明显抑制PHA刺激的淋巴细胞增殖(P<0.05);“第三者”BMSCs可明显抑制双向混合淋巴细胞反应(P<0.05),抑制作用呈BMSCs细胞数量依赖性;隔离腔培养的BMSCs也可明显抑制双向混合淋巴细胞反应(P<0.05)。结论:犬未分化BMSCs在体外具有抑制淋巴细胞增殖的作用,具有一定的调节淋巴细胞反应的能力。
Objective: To study the effect of undifferentiated BMSCs in inhibiting the proliferation of lymphocytes in vitro. Methods: Bone marrow stromal cells were isolated from canine bone marrow and cultured to the second generation, 1 × 105 cells / well BMSCs were stimulated with allogeneic lymphocytes to observe the proliferation of lymphocytes. After stimulating lymphocytes with phytohemagglutinin (PHA, 2ug / mL), allogeneic BMSCs with the number of 1 × 102 ~ 5 cells / well were added to the reaction system to observe the inhibition of lymphocyte proliferation. The number of “third party” BMSCs in the number of 1 × 102 ~ 5 cells / well was added into the mixed lymphocyte reaction system to observe the inhibition of lymphocyte proliferation. Permeability isolation chamber cultured BMSCs were added into two-way mixed lymphocyte reaction system to observe the inhibition of lymphocyte proliferation. Results: BMSCs stimulated allogeneic lymphocytes proliferation in vitro was not obvious; BM-SCs could significantly inhibit PHA-stimulated proliferation of lymphocytes (P <0.05); “Third party” BMSCs could significantly inhibit Biphasic mixed lymphocyte reaction P <0.05). The inhibitory effect was in a dose-dependent manner in BMSCs. BMSCs cultured in isolation chamber also significantly inhibited the two-way mixed lymphocyte reaction (P <0.05). Conclusion: The undifferentiated BMSCs can inhibit the proliferation of lymphocytes in vitro and have certain ability to regulate lymphocyte reaction.