To better characterize murine mesenchymal stem cells (mMSCs), their transcriptome was analyzed via serial analysis of gene expression (SAGE). Abundance of SAGE tags corresponding to interleukin-1 receptor antagonist (IL-1ra) suggested that immunodepleted mMSCs (IDmMSCs) express high levels of these regulatory molecules, which may explain the immunomodulatory activity of MSCs. The goal of this study was to characterize the levels of IL-1ra protein expression in IDmMSCs. mMSCs were isolated from whole bone marrow obtained from the long bones of FVB/N mice. Cells were then suspended in α-MEM media containing L-glutamine, 10% fetal bovine sera, penicillin, and streptomycin and were cultured in a humidified chamber with 5% CO2 for 72 hours at 37°C. MSCs were depleted approximately 7 days later, using antibodies bound to streptavidin-conjugated M-280 Dynabeads. Polymerase chain reaction (PCR) was used to qualitatively determine the presence of IL-1ra, which was confirmed by the presence of the 383 bp product observed. Enzyme-linked immunosorbent assay (ELISA) quantitatively demonstrated that the IDmMSCs produced a high level of IL-1ra protein. Immunostaining revealed that IDmMSCs express high levels of IL-1ra, a natural product thought to counteract the affects of interleukin-1, a proinflammatory cytokine. The results indicate that IL-1ra is expressed in robust levels in subpopulations of the IDmMSCs. Additionally, the present findings implicate that expression of IL1-ra may explain the ability of IDmMSCs to ameliorate the effects of bleomycin-induced lung injury.
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