Selection of Common Markers for Bone Marrow Stromal Cells from Various Bones Using Real-Time RT-PCR: Effects of Passage Number and Donor Age

Abstract

Bone marrow stromal cells (BMSCs) are valuable in tissue engineering and cell therapy, but the quality of the cells is critical for the efficacy of therapy. To test the quality and identity of transplantable cells, we identified the molecular markers that were expressed at higher levels in BMSCs than in fibroblasts. Using numerous BMSC lines from tibia, femur, ilium, and jaw, together with skin and gum fibroblasts, we compared the gene expression profiles of these cells using DNA microarrays and low-density array cards. The differentiation potential of tibia and femur BMSCs was similar to that of iliac BMSCs, and different from jaw BMSCs, but all BMSC lines had many common markers that were expressed at much higher levels in BMSCs than in fibroblasts; several BMSC markers showed discrete expression patterns between jaw and other BMSCs. The common markers are probably useful in routine tests, but their efficacy may depend upon the passage number or donor age. In our study the passage number markedly altered the expression levels of several markers, while donor age had little effect on them. Considering the effects of in vivo location of BMSCs and passage, magnitude of increase in expression levels, and interindividual differences, we identified several reliable markers -- LIF, IGF1, PRG1, MGP, BMP4, CTGF, KCTD12, IGFBP7, TRIB2, and DYNC1I1 -- among many candidates. This marker set may be useful in a routine test for BMSCs in tissue engineering and cell therapy.

Leukemia Inhibitory Factor Secretion is a Predictor and Indicator of Early Progenitor Status in Adult Bone Marrow Stromal Cells

“…Levels of LIF secretion, which was maintained between 86.4 and 166.16 ng=mL per 48 h, reflected this. Our results are similar to those of Igarashi et al, 48 who examined the effect of age on gene expression by iliac crest-derived bone marrow stromal cells. Samples were obtained from seven young donors (aged 18-39) and LIF secretion was compared with that of seven older donors (aged 53-81).…”

Influence of perfusion and cyclic compression on proliferation and differentiation of bone marrow stromal cells in 3-dimensional culture

“…Thus, standard deviations were within a closer range compared with previous experiments (Jagodzinski et al, 2004). Heterogeneous results may be caused by the variation of donor age, passage number, and culture conditions (Siddappa et al, 2007;Igarashi et al, 2007;Bertram et al, 2005). There is still controversy in the literature concerning the effects of culture time.…”

Oleanolic acid exerts an osteoprotective effect in ovariectomy-induced osteoporotic rats and stimulates the osteoblastic differentiation of bone mesenchymal stem cells in vitro

“…Some cytokines that partially participate in osteogenesis were regulated by OA in the early phase, such as the fibroblast growth factor 17 (Fgf17), 18 the fibroblast growth factor (acidic) intracellular binding protein (Fibp), and the insulin-like growth factor (Igf1), the latter of which is also considered to be a reliable marker highly expressed in bMSCs. 19 Moreover, the Notch pathway-related genes, including the anterior pharynx defective 1a homolog (Aph1a), the histone deacetylase 1 (Hdac1), the jagged 1 (Jag1), the numb-like (Numbl), the fringe homolog (Mfng), and the Wnt pathwayYassociated genes, including the mitogen-activated protein kinase kinase kinase 7 interacting protein 1 (Map3k7ip1), responded to OA.…”