Supplementary MaterialsDocument S1
December 16, 2020
Supplementary MaterialsDocument S1. practical microvessels in?vivo, regardless of their anatomical source, or in?situ identity as perivascular or circulating cells. This analysis reveals that muscle pericytes, which are not spontaneously osteochondrogenic as previously claimed, may indeed coincide with an ectopic perivascular subset of committed myogenic cells similar to satellite cells. Cord blood-derived stromal cells, on the other hand, display the unique capacity to form cartilage in?vivo spontaneously, in addition to an assayable osteogenic capacity. These data suggest the need to revise current misconceptions on the origin and function of so-called MSCs, with important applicative implications. The data also support the view that rather than a uniform class of MSCs, different mesoderm derivatives include distinct classes of tissue-specific committed progenitors, possibly of different developmental origin. (Figure?S1A). Dining tables S3 and S2 display the 1st 100 enriched gene models for CB and MU classes, respectively, while Numbers 2A1C2E1 display enrichment heatmaps and plots for selected gene models. The over-represented gene models via gene arranged enrichment evaluation (GSEA) (Subramanian et?al., 2005) support the idea that prospectively purified CB MSCs are extremely proliferative, because the most gene models enriched with this phenotype are linked to proliferation, S stage, DNA and RNA synthesis, or DNA restoration. Alternatively, prospectively purified MU MSCs are obviously seen as a the over-representation of gene XMD8-92 models specifically linked to either muscle development or muscle differentiated function (muscle contraction, muscle development, and energy metabolism). PE and BM expression profiling was analyzed in the same way, but no gene sets were statistically significantly enriched in PE versus CB, BM, and MU, or in BM versus PE, XMD8-92 CB, and MU. However, a number of genes XMD8-92 enriched in BM and PE cells was identified (Table S4). Furthermore, genes associated with hematopoietic support, a defining feature of BM cells, were over-represented in BM cells compared with CB, MU, and PE cells (Figure?S2A). Open in a separate window Figure?2 Enrichment Plots and Heat Maps of Selected Gene Sets for Cord Blood- and Muscle-Derived CD146+ Cells (ACC1) Prenatal CB (cord blood) MSCs are characterized by the over-representation of many genes related to proliferation and cell-cycle regulation. (D and E) Postnatal MU (muscle) MSCs are characterized by the over-representation of tissue-specific genes related to their tissue origin, specifically by genes regulating muscle contraction, muscle development, and energy metabolism. The over-represented gene sets from GSEA showed that prenatal CB CD146+ cells are enriched in gene sets related to proliferation S phase, RNA and DNA synthesis, or DNA repair. For each enriched gene set, the gene expression is also represented as Rabbit polyclonal to Cytokeratin5 a blue-pink ogram in (A1), (B1), and (C1) (CB MSCs), and (D1) and (E1) (MU MSCs). See Supplemental Experimental Procedures for further details of statistical analyses. MSCs from Different Sources Have Radically Different Differentiation Properties BM MSCs, prospectively sorted as CD34?/CD45?/CD146+ and grown under basal conditions that do not induce differentiation, regularly form bone and establish the hematopoietic microenvironment when transplanted heterotopically using an osteoconductive carrier XMD8-92 (Sacchetti et?al., 2007) (Figure?3Aa). Cells sorted based on the same phenotype from BM and other tissues, including MU, were later reported to be highly myogenic both in?vitro and in?vivo, in addition to XMD8-92 sharing the ability to differentiate in culture toward skeletal lineages (Crisan et?al., 2008), based.