目的:通过检测人骨髓间充质干细胞(hMSCs)对共培养的异体树突状细胞活化的细胞因子激活的杀伤细胞(DC-CIK细胞)分泌细胞因子IFN-γ、TNF-α、IL-10、IL-6、IL-4和IL-2的影响,探讨hMSCs的免疫调控机制。方法:从健康人骨髓液中分离、培养hMSCs,通过观察细胞形态,检测其分化为神经元样细胞的神经元烯醇化酶(NSE),脂肪样细胞的油红O染色以及CD29和CD44的表达鉴定hMSCs。从健康人的外周血分离、培养DC,CIK细胞。用流式细胞术(FCM)检测CD1α,HLA-DR的表达来鉴定DC;检测CIK细胞CD3+CD56+的表达。将hMSCs与DC-CIK细胞以1:10的比例共培养4d,用FCM检测培养上清中IFN-γ、TNF-α、IL-10、IL-6、IL-4和IL-2的含量。结果:hMSCs呈长梭形,表达CD29和CD44的阳性细胞数分别为96.6%,94.6%,分化为神经元样细胞的NSE阳性;脂肪样细胞的油红O染色为阳性。DC细胞表达CD1α,HLA-DR的阳性细胞数分别为(91.9±10.04)%,(88.80±8.92)%。DC与CIK细胞共培养第4天表达CD3+CD56+的细胞数为29.23±12.23%。hMSCs与DC-CIK细胞共培养第4天的培养上清中IFN-γ为(135.05±48.19)ng/L;TNF-α为(11.33±1.42)ng/L;IL-10为(10.15±2.25)ng/L;IL-6为(494.63±235.222)ng/L;IL-4为(7.07±2.30)ng/L;IL-2为(1074.63±303.74)ng/L。对照组(DC-CIK细胞)的IFN-γ为(717.60±248.15)ng/L;TNF-α为(17.78±7.52)ng/L;IL-10为(29.95±12.76)ng/L;IL-6为(8.03±0.21)ng/L;IL-4为(9.08±3.07)ng/L;IL-2高于1250ng/L。结论:DC-CIK细胞与hMSCs共培养后,IFN-γ分泌减少,IL-10轻微下调。表明hMSCs可能通过干扰DC-CIK细胞分泌细胞因子来发挥免疫调节作用。 OBJECTIVE: To detect the cytokine IFN-γ, TNF-α and IL-10 secreted by DCs cultured in cocultured allogeneic dendritic cells (DC-CIKs) by detecting human bone marrow mesenchymal stem cells , IL-6, IL-4 and IL-2, to explore the immunomodulatory mechanism of hMSCs. Methods: The hMSCs were isolated and cultured from bone marrow of healthy human. The morphological changes of neuron - like enolase (NSE) and adipose - like cells were observed by observing the cell morphology. The expression of CD29 and CD44 Identification of hMSCs. Isolate from peripheral blood of healthy people and culture DC and CIK cells. The expression of CD1α and HLA-DR was detected by flow cytometry (FCM) to identify DCs. The expression of CD3 + CD56 + in CIK cells was detected. The hMSCs and DC-CIK cells were co-cultured for 1 day at a ratio of 1:10 for 4 days. The contents of IFN-γ, TNF-α, IL-10, IL-6, IL-4 and IL- Results: The hMSCs were spindle-shaped. The number of positive cells expressing CD29 and CD44 were 96.6% and 94.6%, respectively. NSE positive neurons were differentiated into neuron-like cells. Oil-red O staining of adipose-like cells was positive. The positive cells expressing CD1α and HLA-DR in DCs were (91.9 ± 10.04)% and (88.80 ± 8.92)%, respectively. The number of CD3 + CD56 + cells co-cultured with CIK cells on Day 4 was 29.23 ± 12.23%. The levels of IFN-γ in culture supernatants of day 4 were (11.33 ± 1.42) ng / L for TNF-α and (10.15 ± 2.25) for DCs cultured with DC-CIK cells (494.63 ± 235.222) ng / L; IL-4 was (7.07 ± 2.30) ng / L; IL-2 was (1074.63 ± 303.74) ng / L. IL-10 was (29.95 ± 12.76) ng / L, IL-10 was (17.78 ± 7.52) ng / L, IL-10 was (717.60 ± 248.15) ng / 6 was (8.03 ± 0.21) ng / L; IL-4 was (9.08 ± 3.07) ng / L; IL-2 was higher than 1250 ng / L. Conclusion: After co-culture of DC-CIK cells with hMSCs, the secretion of IFN-γ is decreased and IL-10 is slightly down-regulated. HMSCs may play an immunomodulatory role by interfering with the secretion of cytokines by DC-CIK cells.