| Objective:To study the biological characteristics of Flk1~+ mesenchymal stem cells (MSCs) from bone marrow of multiple myeloma patients and to explore its osteogenic potential. To compare the expression of Runx2/Cbfal transcriptional coactivator, TAZ, in the Flk1~+ mesenchymal stem cells from myeloma patients and normal subjects. And to detect the influence of TNF-α and IL-3 on the osteogenic potential and TAZ expression of marrow Flk1~+ MSCs, so as to explore the relationship between TNF-α, IL-3, TAZ and bone disease of multiple myeloma.Methods:Flk1~+ MSCs from bone marrow of multiple myeloma patients were isolated and purified. The morphology of these cells was observed and the growth pattern, immunophonotype, cell cycle and induced differention ability ex vivo were detected. These cells were cultured in the osteogenic medium for two weeks, and real-time RT-PCR was employed to detect the expressions of osteogenic markers before and after osteogenic differention. Von Kossa staining was performed to evaluate the mineral deposition after osteogenic differentiation for two weeks. Real-time RT-PCR and Western-blot were employed to detect mRNA and protein expression of TAZ expression of Flk1~+ MSCs from myeloma patients and normal subjects respectively. The concentrations of TNF-α and IL-3 in the marrow plasma from myeloma patients and normal subjects were detected using ELISA method. CD138~+ myeloma cells or HMCLs were cocultured with normal Flk1~+ MSCs with or without anti-human IL-3 or TNF-α monoclonal antibody in the transwell system. The control group was CD20~+ B lymphocytes from normal subjects. Real-time RT-PCR was employed to detect the mRNA expressions of ALP, Cbfall and TAZ of Flk1~+ mesenchymal stem cells two weeks later. Von Kossa staining was used to detect the mineral deposition. TNF-α and/or IL-3 were added into the culture media of normal...
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