多发性骨髓瘤骨髓间充质干细胞生物学特性分析论文.docVIP

　　多发性骨髓瘤骨髓间充质干细胞生物学特性分析论文 朱步东 任军 王湘漪 李昕 聂鋆 【摘要】 为了研究多发性骨髓瘤（multiple myeloma, MM）病人骨髓间充质干细胞（mesenchymal stem cells, MSC）的病理生物学特性，以探讨MSC在MM骨损伤发生中的作用，取11例初治MM病人和5例正常人骨髓，用贴壁法体外分离培养MSC 。应用流式细胞术分析MM骨髓MSC表型，MTT法检测MSC增殖能力，组织化学染色测定细胞向成骨细胞和脂肪细胞分化能力。应用酶联免疫吸附实验（enzyme-linked immunosorbent assay, ELISA）测定细胞培养上清液中白介素-6（interleukin-6，IL-6）和干细胞生长因子（stem cell factor, SCF）浓度。收集MSC培养上清作为条件培养液，按梯度浓度加入人SKO007骨髓瘤细胞系培养体系中.freelal Stem Cells Derived from Bone Marroa AbstractThe study esenchymal stem cells (MSCs) in the pathogenesis of bone disease particularly observed in multiple myeloma (MM), the biological features of marro patients 11 neal adults and MSCs etric technique. The proliferation of MSCs ical staining. The concentrations of IL-6 and SCF in the culture supernatant e-linked immunosorbent assay (ELISA). MSC culture supernatants ed to evaluate their support on the proliferation of an MM cell line SKO007 cells. The results shoarro MM patients ogenously positive for CD29, CD73, CD166 and HLA-ABC and negative for hematopoietic cell marker CD45 and endothelial cell marker CD31, the phenotype of ilar to that of marro normal adults. MTT assay indicated that MSCs from MM patients or normal adults proliferated at similar rates. MSCs from MM patients occupied in vitro osteogenic and adipogenic capacity as those from normal adults. The levels of IL-6 and SCF in culture supernatant ore, MSC culture supernatants from MM bone marroote the proliferation of SKO007 cells. It is concluded that marro bone marroal in their proliferation and differentiation capacities, and myeloma bone disease may not be ascribed to the differentiation of MSCs ay provide necessary cues for the survival of malignant myeloma cells. Key esenchymal stem cell; multiple myeloma; cytokine 多发性骨髓瘤（multiple myeloma, MM）患者呈现特征性的骨质破坏。这种骨质损伤与破骨细胞数量增加，成骨细胞数减少，从而导致骨吸收与成骨之间失平衡有关［1］。间充质干细胞（mesenchymal stem cells,.freel Cells公司产品），培养48小时后去除非贴壁细胞，继续培养至贴壁细胞约80%融合，收集培养上清于-70℃冻存备用。0.05%胰蛋白酶消化传代培养。第3代细胞用于以下实验。 细胞免疫表型分析 胰蛋白酶消化收集MSC，