Sa Ya-Lian, Shen Xiao-Mei, Shi Ke-Qian, Tang Hui, Zhao Ren-Bin, Lu Jie, Song Jian-Xin, Yan Xin-Min
Institute of Clinical and Basic Medical Sciences; The First People's Hospital of Yunnan Province (Kunhua Hospital Affiliated to Kunming Medical University), Kunming 650032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2010 Oct;26(10):988-91.
to study the effect of human bone marrow derived mesenchymal stem cells (hMSCs) on cytokines secretion (IFN-γ, TNF-α, IL-10, IL-6, IL-4 and IL-2) of allogeneic DC-CIK cells (in co-culture of CIK cells with DC), which investigate the mechanism of immunoregulation induced by hMSCs.
the hMSCs from bone marrow were isolated, expanded and identified by cell morphology, differentiation into neuron-like cells with NSE, fat-like cells with red-oil stain, and expression of CD29, CD44. The DC and CIK cells from peripheral blood were isolated, expanded and identified by CD1α, HLA-DR or CD3(+);CD56(+);. The hMSCs were co-cultured with DC-CIK cells according to ratio 1:10. The expression of the six cytokines in supernatant was evaluated by flow cytometry after 4 days of DC-activated CIK cells in co-culture with hMSCs.
the hMSCs displayed a fibroblast-like morphology and the positive cells of CD29 and CD44 were 96.6%, 94.6%, which have the capacity of differentiation into neuron-like cells with expressed NSE as well as fat-like cells with red-oil stain positive. The expression of CD1α, HLA-DR in DC was (91.9 ± 10.04)% and (88.8 ± 8.92)%. The CD3(+);CD56(+); double positive cells in DC-CIK cells was (29.23 ± 12.23)% compared to CIK cells with (15.98 ± 2.49)%. The cytokines secretion of DC-CIK cells in co-culture with hMSCs was IFN-γ (135.05 ± 48.19) ng/L; TNF-α (11.33 ± 1.42) ng/L; IL-10 (10.15 ± 2.25) ng/L; IL-6 (494.63 ± 235.222) ng/L; IL-4 (7.07 ± 2.30) ng/L and IL-2 (1074.6 3 ± 303.74) ng/L. In control group (DC-CIK cells) the secretion of IFN-γ, TNF-α, IL-10, IL-6, IL-4 and IL-2 was (717.6 ± 248.15) ng/L; (17.78 ± 7.52) ng/L; (29.95 ± 12.76) ng/L; (8.03 ± 0.21) ng/L, (9.08 ± 3.07) ng/L as well as IL-2 1 250 ng/L.
the secretion of IFN-γ and IL-10 were down-regulated. It probably implied that hMSCs had the effect of immunoregulation on DC-CIK cells.
研究人骨髓间充质干细胞（hMSCs）对异基因DC-CIK细胞（CIK细胞与DC共培养）细胞因子分泌（IFN-γ、TNF-α、IL-10、IL-6、IL-4和IL-2）的影响，探讨hMSCs诱导免疫调节的机制。
从骨髓中分离、扩增hMSCs，并通过细胞形态、分化为表达神经元特异性烯醇化酶（NSE）的神经元样细胞、用油红染色分化为脂肪样细胞以及CD29、CD44的表达进行鉴定。从外周血中分离、扩增DC和CIK细胞，并通过CD1α、HLA-DR或CD3(+)；CD56(+)进行鉴定。hMSCs与DC-CIK细胞按1:10的比例共培养。在hMSCs与DC激活的CIK细胞共培养4天后，通过流式细胞术评估上清液中六种细胞因子的表达。
hMSCs呈现成纤维细胞样形态，CD29和CD44阳性细胞分别为96.6%、94.6%，具有分化为表达NSE的神经元样细胞以及油红染色阳性的脂肪样细胞的能力。DC中CD1α、HLA-DR的表达分别为(91.9±10.04)%和(88.8±8.92)%。与CIK细胞（15.98±2.49）%相比，DC-CIK细胞中CD3(+)；CD56(+)双阳性细胞为(29.23±12.23)%。与hMSCs共培养的DC-CIK细胞的细胞因子分泌情况为：IFN-γ（135.05±48.19）ng/L；TNF-α（11.33±1.42）ng/L；IL-10（10.±）ng/L；IL-6（494.63±235.222）ng/L；IL-4（7.07±2.30）ng/L；IL-2（1074.63±30±）ng/L。对照组（DC-CIK细胞）中IFN-γ、TNF-α、IL-10、IL-6、IL-4和IL-2的分泌分别为（717.6±248.15）ng/L；（17.78±7.52）ng/L；（±）ng/L；（8.03±0.21）ng/L，（9.08±3.07）ng/L以及IL-2 1250 ng/L。
IFN-γ和IL-10的分泌下调。这可能意味着hMSCs对DC-CIK细胞具有免疫调节作用。
