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目的:研究犬脂肪基质干细胞(adipose derived stem cells,ADSCs)体外抑制淋巴细胞增殖的作用。方法:从Beagle犬腹股沟脂肪组织中分离获取脂肪基质干细胞,体外培养至第二代,1×105个/孔ADSCs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,2μg/mL)刺激淋巴细胞后,分别以1×102～5细胞/孔数量的异体ADSCs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”ADSCs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ADSCs加入混合双向淋巴细胞反应体系中,观察淋巴细胞增殖抑制情况。结果:ADSCs刺激同种异体淋巴细胞体外增殖作用不明显;ADSCs可明显抑制PHA刺激的淋巴细胞增殖(P<0.05);“第三者”ADSCs可明显抑制双向混合淋巴细胞反应(P<0.05),抑制作用呈ADSCs细胞数量依赖性;隔离腔培养的ADSCs也可明显抑制双向混合淋巴细胞反应(P<0.05)。结论:ADSCs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 inhibitory effect of ADSCs on the proliferation of lymphocytes in vitro. METHODS: Adipose-derived stem cells were isolated from the adipose tissue of the inguinal groin of Beagle dogs and cultured to the second passage. The cells were stimulated with 1 × 105 cells / well of ADSCs for allogeneic lymphocytes to observe the proliferation of lymphocytes. After stimulating lymphocytes with phytohemagglutinin (PHA, 2μg / mL), allogeneic ADSCs 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” ADSCs with the number of 1 × 102 ~ 5 cells / well was added to the two-way mixed lymphocyte reaction system to observe the inhibition of lymphocyte proliferation. Permeability isolation chamber cultured ADSCs were added to the mixed bi-directional lymphocyte reaction system to observe the inhibition of lymphocyte proliferation. Results: ADSCs stimulated allogeneic lymphocytes proliferation in vitro was not obvious; ADSCs could significantly inhibit PHA-stimulated lymphocyte proliferation (P <0.05); “third party” ADSCs could significantly inhibit bidirectional mixed lymphocyte reaction (P < 0.05). The inhibitory effect of ADSCs in ADSCs was dependent on the number of ADSCs. Isolation of cultured ADSCs significantly inhibited the two-way mixed lymphocyte reaction (P <0.05). Conclusion: ADSCs can inhibit the proliferation of lymphocytes in vitro and have certain ability of regulating lymphocyte reaction.