|Application:||Gene expression microarray analysis|
|Number of samples:||8|
|Release date:||Apr 2 2009|
|Last update date:||Jan 18 2013|
|Diseases:||Autoimmune Diseases, Diabetes Mellitus, Type 1, Hyperglycemia, Intestinal Pseudo-Obstruction, Neoplasms|
|Dataset link||Immunomodulatory function of bone marrow-derived mesenchymal stem cells in experimental autoimmune type 1 diabetes|
To generate MSC, BM mononuclear cells were isolated from the femurs and tibiae of at least 5 mice in order to minimize cell variability. Cells are seeded in flasks at a concentration of 10x106/25 cm2 in M10 medium (DMEM medium [Cambrex, East Rutherford, New Jersey] containing 10% fetal calf serum [HyClone, Logan, Utah], 1% penicillin-streptomycin, and 1% L-glutamine [both from Cambrex]). To examine MSC in an inflammatory setting, 7.5x105 NOD- or BALB/c-MSC/well were cultured for 48h in 6-well plates in M10 medium containing 10 ng/ml recombinant murine IL-1beta (Peprotech, Rocky Hill, NJ). To ensure that our cultured cells had multipotent potential, we tested MSC P4 cultures for their ability to undergo differentiation into chondrocytes, osteocytes, and adipocytes as previously published.10 Chondrogenic differentiation was induced by 50 ug/ml ascorbic acid and 1 ng/ml TGF?-1 (Peprotech), osteogenic differentiation was induced by 50 ug/ml ascorbic acid, 10 mM sodium alpha-glycerophosphate, and 10-8 M dexamethasone, and adipogenic differentiation was induced by 10-7 M dexamethasone and 6 ng/ml insulin. MSC were analyzed for expression at passage 4 (P4). We examined the gene expression profile of NOD- (samples #1 to 4) and BALB/c-MSC (samples #5 to 8), obtained from normoglycemic NOD mice and age-matched BALB/c mice. A striking concordance in the pattern of gene regulation across the entire screen was observed when BALB/c-MSC genes were compared to the NOD-MSC 4 different culture of NOD derived MSC were compared to 4 differente cell culture of Balb/c derived MSC, Balb/c cell coltures were considered reference samples.