A simple and reliable protocol for long-term culture of murine bone marrow stromal (mesenchymal) stem cells that retained their in vitro and in vivo stemness in long-term culture

Table 1 Upregulated osteoblastic genes by BMSCs-FS (p25) versus ST2 cells

Gene name	Gene symbol	Fold
Ossification and matrix molecules
Alkaline phosphatase, liver/bone/kidney	Alpl	7.9
Bone gamma carboxyglutamate protein	Bglap	2.1
Collagen, type I, alpha 1	Col1a1	7.5
Collagen, type I, alpha 2	Col1a2	4.2
Fibroblast growth factor receptor 1	Fgfr1	4.4
Fibroblast growth factor receptor 2	Fgfr2	2.3
Insulin-like growth factor I receptor	Igf1r	2.2
BMP signaling pathway
Bone morphogenetic protein 2	Bmp2	74
Bone morphogenetic protein 3	Bmp3	25.3
Bone morphogenetic protein 5	Bmp5	2
Bone morphogenetic protein 6	Bmp6	17.7
Bone morphogenetic protein receptor, type 1B	Bmpr1b	3.8
Bone morphogenetic protein 4	Bmp4	2
Extracellular Matrix (ECM) Molecules
Biglycan	Bgn	2
Collagen, type II, alpha 1	Col2a1	7.5
Collagen, type III, alpha 1	Col3a1	2.2
Collagen, type XIV, alpha 1	Col14a1	2.2
Collagen, type V, alpha 1	Col5a1	2.6
Integrin alpha 2	Itga2	10
Integrin alpha 2b	Itga2b	1.5
Matrix metallopeptidase 2	Mmp2	17
Matrix metallopeptidase 8	Mmp8	12
Matrix metallopeptidase 9	Mmp9	23.7
Osteogenic growth factors
Colony stimulating factor 1 (macrophage)	Csf1	3
Fibroblast growth factor 1	Fgf1	2.5
Fibroblast growth factor 2	Fgf2	2.7
Growth differentiation factor 10	Gdf10	3.7
Insulin-like growth factor 1	Igf1	2
Platelet derived growth factor, alpha	Pdgfa	3.2
Osteogenic transcription factors
Runt related transcription factor 2	Runx2	4.5
Sp7 transcription factor 7	Sp7	6.7

Cells were induced into osteoblast differentiation as described in the M&M. Mouse osteogenesis RT² Profiler™ PCR array with 84 osteogenic genes was performed using the SYBR® Green quantitative PCR method. Each target gene was normalized to a group of reference genes as described by manufacture instruction’s. Up-regulated genes (≥ 2 fold) by BMSCs-FS are represented as fold change over ST2 cells

ISSN: 1480-9222