Molecular Analysis of the Differentiation Potential of Murine Mesenchymal Stem Cells from Tissues of Endodermal or Mesodermal Origin

Abstract

Mesenchymal stem cells (MSCs) have received great attention due to their remarkable regenerative, angiogenic, antiapoptotic, and immunosuppressive properties. Although conventionally isolated from the bone marrow, they are known to exist in all tissues and organs, raising the question on whether they are identical cell populations or have important differences at the molecular level. To better understand the relationship between MSCs residing in different tissues, we analyzed the expression of genes related to pluripotency (SOX2 and OCT-4) and to adipogenic (C/EBP and ADIPOR1), osteogenic (OMD and ALP), and chondrogenic (COL10A1 and TRPV4) differentiation in cultures derived from murine endodermal (lung) and mesodermal (adipose) tissue maintained in different conditions. MSCs were isolated from lungs (L-MSCs) and inguinal adipose tissue (AMSCs) and cultured in normal conditions, in overconfluence or in inductive medium for osteogenic, adipogenic, or chondrogenic differentiation. Cultures were characterized for morphology, immunophenotype, and by quantitative real-time reverse transcription-polymerase chain reaction for expression of pluripotency genes or markers of differentiation. Bone marrow-derived MSCs were also analyzed for comparison of these parameters. L-MSCs and A-MSCs exhibited the typical morphology, immunophenotype, and proliferation and differentiation pattern of MSCs. The analysis of gene expression showed a higher potential of adipose tissue-derived MSCs toward the osteogenic pathway and of lung-derived MSCs to chondrogenic differentiation, representing an important contribution for the definition of the type of cell to be used in clinical trials of cell therapy and tissue engineering.

Cardiac Fibroblasts Adopt Osteogenic Fates and Can Be Targeted to Attenuate Pathological Heart Calcification

“…In contrast, genes that were minimally expressed in cardiac fibroblasts were induced in a specific temporal manner during the course of osteogenic differentiation (Figure 1C) and included sets of genes known to regulate inflammation, extracellular matrix proteins and cell metabolism (Figure S1A). Next, we created an osteogenic signature based on a set of 37 genes that are induced during osteogenic differentiation (Chen et al, 2012);(Choi et al, 2010);(Graneli et al, 2014);(Harkness et al, 2011);(Hoshiba et al, 2009);(Liu et al, 2013);(Miguez et al, 2014);(Nora et al, 2012);(Olivares-Navarrete et al, 2011) (Figure S1B). We used the mean fold change in expression of this set of genes to quantitatively determine an osteogenic signature and observed that compared to control cardiac fibroblasts, cardiac fibroblasts subjected to osteogenic differentiation progressively adopted an osteogenic signature (Figure 1D).…”

“…1685 genes with a FDR < 0.05 and a minimum 2 fold (log 2 ) change were subsequently clustered and visualized. A set of osteogenic genes known to be induced during bone formation or osteogenic differentiation was compiled from the literature to determine an osteogenic signature (Chen et al, 2012); (Choi et al, 2010); (Graneli et al, 2014); (Harkness et al, 2011); (Hoshiba et al, 2009); (Liu et al, 2013); (Miguez et al, 2014); (Nora et al, 2012); (Olivares-Navarrete et al, 2011). All RNA-seq analysis were performed with ≥ 2 biological replicates.…”

Differences in Gene Expression and Cytokine Release Profiles Highlight the Heterogeneity of Distinct Subsets of Adipose Tissue-Derived Stem Cells in the Subcutaneous and Visceral Adipose Tissue in Humans

“…gene expression changes during in vitro differentiation to osteogenic, chondrogenic or adipogenic lineages [30]; [31], ii.) comparative analysis of bone marrow and adipose tissue-derived mesenchymal stromal cells in in vitro adipogenic, chondrogenic or osteogenic differentiation [32]; [33], and iii.)…”

Comparison of endometrial regenerative cells and bone marrow stromal cells

“…These studies have found that stromal cells produced from these various cell and tissue types have a similar phenotype [28-32]. All express CD44, CD73, CD90 and CD105, but do not express hematopoietic cell markers.…”