摘要
背景与目的:我们先前的实验证明,在适宜的刺激下骨髓间充质干细胞(mesenchymal stem cells,MSCs)可分泌可溶性的细胞因子。本实验旨在研究MSCs对慢性粒细胞白血病(chronic myeloid leukemia,CML)细胞增殖的影响,并检测上清液中MSCs分泌的细胞因子。方法:采集健康供者骨髓培养MSCs,流式细胞仪检测第三代MSCs表面标记。将MSCs与不同浓度的CML患者的骨髓单个核细胞(chronic myeloid leukemia mononuclear cells,CML-MNC)共培养;每天收集各组CML-MNC计数取均值,绘制生长曲线。于实验的第3天和第6天收集各培养组的上清液,ELISA法测IFN-α。结果:原代和传代培养的细胞呈大的长梭形,增殖旺盛;细胞表面标记:CD44阳性,而CD45阴性。生长曲线示:与MSCs培养的CML-MNC增殖受抑。ELISA法检测结果示:对照组CML细胞生成IFN-α极低,显著低于各实验组,差异有统计学意义(P<0.001)。MSCs与CML细胞共培养时可以分泌大量的IFN-α,且随着MSCs浓度的增加和共培养时间的延长,生成的IFN-α量增多。结论:MSCs与CML细胞共培养时能产生大量IFN-α,抑制CML细胞增殖。
Background and Objective：Bone marrow mesenchymal stem cells （MSCs） are of particular interest due their potential clinical use in tissue engineering. Previous studies showed that MSCs could secret soluble factor （s） upon stimulation. This study was to evaluate the influence of human bone marrow MSCs on proliferation of chronic myeloid leukemia cells, and assess the secretion of cytokines in the supernatant by MSCs. Methods： Bone marrow MSCs extracted from healthy donors were cultured in DMEM-LG. The surface markers on the third passage MSCs were detected by flow cytometry. MSCs were co-cultured with chronic myeloid leukemia mononuclear cells （CML- MNCs） at various concentrations. Ceil proliferation was measured by flow cytometry. The interferon （IFN）-α level in the supernatant was analyzed by ELISA assay. Results; The primary and passaged MSCs mostly appeared fibroblast-like and showed strong capacity of growth and reproduction. The membrane marker CD44 was positive and CD45 was negative on the surface of MSCs. Co-culture of MSCs with CML-MNC significantly inhibited the proliferation of CML-MNC. The IFN-α level in the supernatant of cell culture was significantly higher in the co-culture group than in the CML-MNC control group （P〈0.001）. Secretion of IFN-αwas elevated with the increase of the MSC concentration and co-culture duration. Conclusion： Co-culture of MSCs with CML could secrete a substantial amount of IFN-α, thus to inhibit the proliferation of CML.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2009年第1期39-42,共4页
Chinese Journal of Cancer
关键词
骨髓
间充质干细胞
粒细胞白血病
干扰素-Α
细胞增殖
bone marrow, mesenchymal stem cells, myeloid leukemia,interferon （IFN）-α, cell proliferation
