Project name: Homo sapiens

Description: Mesenchymal stem/stromal cells (MSCs) derived from the BM of healthy donors (dMSCs) and myeloma patients (pMSCs) were co-cultured with the model myeloma cell line - MM.1S -, and the gene expression profile of MSCs induced by this interaction was analyzed using high density oligonucleotide microarrays. Deregulated genes in co-culture common to both d/pMSCs revealed functional involvement in tumor microenvironment cross-talk, myeloma growth induction and drug resistance, angiogenesis and signals for osteoclast activation and inhibition of osteoblasts. Additional genes induced by co-culture were exclusively deregulated in pMSCs and were predominantly associated to RNA processing, the ubiquitine-proteasome pathway, regulation of cell cycle and Wnt signaling.Overall design: Primary MSCs from bone marrow (BM) samples of healthy donors (n=8) and multiple myeloma (MM) patients (n=13) were generated as described by Garayoa et al. 2009 (PMID:19357701). Ficoll-Paque density gradient separation medium was used to isolate mononuclear cells from BM aspirates. MSCs were selected by adherence to plastic and subsequently expanded until passage 2 when they were used for co-culture. For the MSC-MM.1S co-cultures we used a 6-well format transwell system with 1 µm pore size membrane. Briefly, 2 X 10^4 MSCs were first cultured attached to the lower side of the membrane, and when reached ≈ 85% confluence 1 X 10^6 MM.1S cells were seeded on the upper side. After 24 h co-culture in RPMI 1640 medium supplemented with 10% FBS and antibiotics, MSCs were recovered by trypsinization, washed once in PBS and stored at -80°C in RLT buffer (Qiagen) until RNA extraction. Absence of MM.1S cells in the recovered MSC population was assessed in selected samples by flow cytometry analysis.

Data type: Transcriptome or Gene expression

Sample scope: Multiisolate

Relevance: Medical

Last updated: 2013-04-15