源于间充质干细胞的神经细胞抑制异体T淋巴细胞增生的实验研究

Proliferation inhibition of allogenic T lymphocytes by neurons drived from mesenchymal stem cells

目的研究来源于骨髓间充质干细胞(MSCs)的神经细胞体外调节T淋巴细胞增殖的功能。方法从人骨髓中分离培养MSCs,将MSCs向神经细胞诱导分化1￣2周。根据诱导分化的时间分为:诱导分化前组、诱导分化1周组和诱导分化2周组。以1×104/孔不同诱导分化时间的细胞作为刺激细胞,分离纯化的异体T淋巴细胞作为反应细胞,共培养6d后,检测T淋巴细胞增殖能力。植物血凝素(PHA)刺激T淋巴细胞后,分别与各组细胞共培养,观察各组细胞对非特异性T淋巴细胞增殖的作用。为模拟体内炎症环境,各组细胞经γ-IFN预处理48h后,再与T淋巴细胞混合培养,观察T淋巴细胞增殖被抑制的情况。结果MSCs向神经细胞分化后抑制T淋巴细胞增殖,且可显著抑制PHA刺激的非特异性T淋巴细胞增殖。γ-IFN预处理使抑制作用进一步加强。抑制作用随着MSCs向神经细胞分化,呈诱导分化的时间依赖性增强。结论源于MSCs的神经细胞在体外抑制T淋巴细胞增殖反应,为细胞移植治疗神经系统损伤提供了试验依据。

Objective To study whether neurocytes which were induced from bone marrow mesenchymal stem cells （MSCs） can suppress allogenic T lymphocyte proliferation, and explore its possible mechanisms. Methods Human MSCs were isolated from bone marrow, and cultured and induced to differentiate into neurocytes for 1-2 weeks. According to the differentiation time, MSCs were divided into 3 groups： undifferentiated group, l-week differentiated group and 2-week differentiated group. The cells at 1×10^4 /well were co-cultured with allogenic T lymphocytes for 6 d. T lymphocyte proliferation was assessed by [^3H]-thymidine incorporation using a liquid scintillation counter. To study the effect of the cells of each group modulated non-specific T lymphocyte proliferation, PHA （2 μg/mL） were added into coculture medium as a stimulate factor before the cells of each group were added. To simulate the inflammation environment in vivo, befor coculture with T lymphocytes, the cells of each group were pretreated by γ-IFN for 48 h. Results Neurocytes fxom MSCs could suppress the proliferation of allogenic T lymphocytes, including PHA-stimulated non-specific T lymphocytes. Pretreated by γ-IFN, the cells had a stronger ability of inhibiting the T lymphocyte proliferation. The proliferation-inhibiting effect was increased in a induced-differentiation time dependent manner. Conclusion Neurocytes fxom bone marrow MSCs can suppress the proliferation of allogenic T lymphocytes in vitro, which demonstrates that the immunity property of differented MSCs into neurocytes offers a new possibility in the development of cell therapies for neurological diseases.