Phenotype, donor age and gender affect function of human bone marrow derived mesenchymal stromal cells
BackgroundMesenchymal stromal cells (MSCs) are attractive for cell based therapies ranging from regenerative medicine and tissue engineering to immunomodulation. However, clinical efficacy is variable and it is unclear how the phenotypes defining bone marrow (BM) derived MSCs as well as donor characteristics affect their functional properties.MethodsBM MSCs were isolated from 53 (25 female, 28 male; age: 13 to 80 years) donors and analyzed by: (1) phenotype using flow cytometry and cell size measurement; (2) in vitro growth kinetics using population doubling time; (3) colony formation capacity and telomerase activity; and (4) function by in vitro differentiation capacity, suppression of T cell proliferation, cytokines and trophic factors secretion, and hormone and growth factor receptor expression. Additionally, expression of Oct4, Nanog, Prdm14 and SOX2 mRNA was compared to pluripotent stem cells.ResultsBM MSCs from younger donors showed increased expression of MCAM, VCAM 1, ALCAM, PDGFRb, PDL 1, Thy1 and CD71, and led to lower IL 6 production when co cultured with activated T cells. Female BM MSCs showed increased expression of IFN gR1 and IL 6b, and were more potent in T cell proliferation suppression. High clonogenic BM MSCs were smaller, divided more rapidly and were more frequent in BM MSC preparations from younger female donors. CD10, b1integrin, HCAM, CD71, VCAM 1, IFN gR1, MCAM, ALCAM, LNGFR and HLA ABC were correlated to BM MSC preparations with high clonogenic potential and expression of IFN gR1, MCAM and HLA ABC was associated with rapid growth of BM MSCs. The mesodermal differentiation capacity of BM MSCs was unaffected by donor age or gender but was affected by phenotype (CD10, IFN gR1, GD2) BM MSCs from female and male donors expressed androgen receptor and FGFR3, and secreted VEGF A, HGF, LIF, Angiopoietin 1, basic fibroblast growth factor (bFGF) and NGFB. HGF secretion correlated negatively to the expression of CD71, CD140b and Galectin 1. The expression of Oct4, Nanog and Prdm14 mRNA in BM MSCs was much lower compared to pluripotent stem cells and was not related to donor age or gender. Prdm14 mRNA expression correlated positively to the clonogenic potential of BM MSCs.ConclusionsBy identifying donor related effects and assigning phenotypes of BM MSC preparations to functional properties, we provide useful tools for assay development and production for clinical applications of BM MSC preparations.
BMC Medicine