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[多发性骨髓瘤患者骨髓来源间充质干细胞的生物学特性]

[Biological properties of mesenchymal stem cells derived from bone marrow of patients with multiple myeloma].

作者信息

Zhu Bu-Dong, Ren Jun, Wang Xiang-Yi, Li Xin, Nie Jun

机构信息

Department of Oncology, Peking University School of Oncology & Beijing Cancer Hospital, Beijing Institute for Cancer Research, Beijing 100036, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2006 Dec;14(6):1138-42.

Abstract

The study was purposed to explore the role of mesenchymal stem cells (MSCs) in the pathogenesis of bone disease particularly observed in multiple myeloma (MM), the biological features of marrow derived MSCs from patients with MM have been investigated. Marrow aspirates were harvested from 11 newly diagnosed patients with MM and 5 normal adults and MSCs were isolated and culture-expanded by the cell properties of adherence to plastic flasks, The phenotype was analyzed by flow cytometric technique. The proliferation of MSCs was observed by MTT assay and their differentiation capacities into osteoblasts and adipoblasts were assessed with lineage-specific histochemical staining. The concentrations of IL-6 and SCF in the culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA). MSC culture supernatants were collected and MTT assay was performed to evaluate their support on the proliferation of an MM cell line SKO007 cells. The results showed that bone marrow-derived MSCs from MM patients were homogeneously positive for CD29, CD73, CD166 and HLA-ABC and negative for hematopoietic cell marker CD45 and endothelial cell marker CD31, the phenotype of which was similar to that of marrow counterparts from 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 were greatly up-regulated in MM patients by ELISA assay. Furthermore, MSC culture supernatants from MM bone marrow displayed enhanced activity to promote the proliferation of SKO007 cells. It is concluded that marrow-derived MSCs from bone marrow of MM patients are normal in their proliferation and differentiation capacities, and myeloma bone disease may not be ascribed to the differentiation of MSCs while the elevated secretion of IL-6 and SCF may provide necessary cues for the survival of malignant myeloma cells.

摘要

本研究旨在探讨间充质干细胞(MSCs)在骨病发病机制中的作用,尤其是在多发性骨髓瘤(MM)中观察到的骨病,对MM患者骨髓来源的MSCs的生物学特性进行了研究。从11例新诊断的MM患者和5例正常成年人中采集骨髓抽吸物,根据其贴壁于塑料培养瓶的细胞特性分离并培养扩增MSCs,采用流式细胞术分析其表型。通过MTT法观察MSCs的增殖情况,并用谱系特异性组织化学染色评估其向成骨细胞和脂肪细胞的分化能力。采用酶联免疫吸附测定(ELISA)检测培养上清液中IL-6和SCF的浓度。收集MSCs培养上清液,进行MTT法检测以评估其对MM细胞系SKO007细胞增殖的支持作用。结果显示,MM患者骨髓来源的MSCs对CD29、CD73、CD166和HLA-ABC呈均匀阳性,对造血细胞标志物CD45和内皮细胞标志物CD31呈阴性,其表型与正常成年人骨髓来源的MSCs相似。MTT法表明,MM患者或正常成年人的MSCs增殖速率相似。MM患者的MSCs与正常成年人的MSCs在体外具有相同的成骨和成脂能力。ELISA检测显示,MM患者培养上清液中IL-6和SCF的水平显著上调。此外,MM骨髓来源的MSCs培养上清液促进SKO007细胞增殖的活性增强。结论是,MM患者骨髓来源的MSCs在增殖和分化能力方面是正常的,骨髓瘤骨病可能不归因于MSCs的分化,而IL-6和SCF分泌的增加可能为恶性骨髓瘤细胞的存活提供必要条件。

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