Molecular profile and cellular characterization of human bone marrow mesenchymal stem cells: Donor influence on chondrogenesis

Abstract

a b s t r a c tBackground: The use of autologous or allogenic stem cells has recently been suggested as an alternative therapeutic approach for treatment of cartilage defects. Bone marrow mesenchymal stem cells (BM-MSCs) are well-characterized multipotent cells that can differentiate into different cell types. Understanding the potential of these cells and the molecular mechanisms underlying their differentiation should lead to innovative protocols for clinical applications. The aim of this study was to evaluate the usefulness of surface antigen selection of BM-MSCs and to understand the mechanisms underlying their differentiation. Methods: MSCs were isolated from BM stroma and expanded. CD105 + subpopulation was isolated using a magnetic separator. We compared culture-expanded selected cells with non-selected cells. We analyzed the phenotypic profiles, the expression of the stem cell marker genes Nanog, Oct3/4, and Sox2 and the multi-lineage differentiation potential (adipogenic, osteogenic, and chondrogenic). The multilineage differentiation was confirmed using histochemistry, immunohistochemistry and/or real-time polymerase chain reaction (qPCR) techniques. Results: The selected and non-selected cells displayed similar phenotypes and multi-lineage differentiation potentials. Analyzing each cell source individually, we could divide the six donors into two groups: one with a high percentage of CD29 (b1-integrin) expression (HL); one with a low percentage of CD29 (LL). These two groups had different chondrogenic capacities and different expression levels of the stem cell marker genes. Conclusions: This study showed that phenotypic profiles of donors were related to the chondrogenic potential of human BM-MSCs. The chondrogenic potential of donors was related to CD29 expression levels. The high expression of CD29 antigen seemed necessary for chondrogenic differentiation. Further investigation into the mechanisms responsible for these differences in BM-MSCs chondrogenesis is therefore warranted. Understanding the mechanisms for these differences will contribute to improved clinical use of autologous human BM-MSCs for articular cartilage repair.

Surface Markers for Chondrogenic Determination: A Highlight of Synovium-Derived Stem Cells

“…After incubation in chondrogenic induction medium [chemically defined Dulbecco’s Modified Eagle Medium (DMEM) with 1 ng/mL of recombinant human TGF-β3], the high CD29 expression group displayed a greater chondrogenic differentiation capacity than the low CD29 expression group, while the low expression subpopulation showed no difference compared to the control (DMEM with 20% fetal bovine serum). This result was verified with immunohistochemical analysis of type II collagen, as well as RT-PCR analysis of chondrocyte-specific genes [40]. The relationship between chondrogenic potential and CD29 expression is evident in BMSCs; however, the relationship with CD29 and SDSCs remains uncertain.…”

Enhanced chondrogenesis of bone marrow-derived stem cells by using a combinatory cell therapy strategy with BMP-2/TGF-β1, hypoxia, and COL1A1/HtrA1 siRNAs

“…Unfortunately, few studies have examined the benefits of cell selection criteria for cartilage engineering applications. Whereas CD105 enrichment using a magnetic separator seemed not to influence the multilineage potential of BM-MSCs, the high expression of CD29 seemed necessary for chondrogenic differentiation 44 . Others found that CD29 + /CD90 + selected MSCs from rat bone marrow displayed a significantly reduced capacity for osteogenic/adipogenic differentiation 45 .…”

The effect of cyclic hydrostatic pressure on the functional development of cartilaginous tissues engineered using bone marrow derived mesenchymal stem cells

“…The finding in this study that the response to loading is donor dependent may in part explain some of this variability. Perhaps this result is to be expected given the well documented variability in the response of MSCs from different donors to cytokine induced chonodrogenic differentiation (Cicione et al 2010), and the fact that animal model studies investigating mechanically induced chondrogenesis in vivo often report dramatic donor dependant response to loading (Khayyeri et al ;Tägil and Aspenberg 1999;De Rooij et al 2001). Further studies are required to better understand donor variability in the response to mechanical signals that clearly cannot be determined based on an analysis of only a small number of donors.…”