Single-Cell Transcriptomic Analysis Reveals the Cellular Heterogeneity of Mesenchymal Stem Cells

Abstract

Ex vivo-expanded mesenchymal stem cells (MSCs) have been demonstrated to be a heterogeneous mixture of cells exhibiting varying proliferative, multipotential, and immunomodulatory capacities. However, the exact characteristics of MSCs remain largely unknown. By single-cell RNA sequencing of 61,296 MSCs derived from bone marrow and Wharton’s jelly, we revealed five distinct subpopulations. The developmental trajectory of these five MSC subpopulations was mapped, revealing a differentiation path from stem-like active proliferative cells (APCs) to multipotent progenitor cells, followed by branching into two paths: 1) unipotent preadipocytes or 2) bipotent prechondro-osteoblasts that were subsequently differentiated into unipotent prechondrocytes. The stem-like APCs, expressing the perivascular mesodermal progenitor markers CSPG4/MCAM/NES, uniquely exhibited strong proliferation and stemness signatures. Remarkably, the prechondrocyte subpopulation specifically expressed immunomodulatory genes and was able to suppress activated CD3+ T cell proliferation in vitro, supporting the role of this population in immunoregulation. In summary, our analysis mapped the heterogeneous subpopulations of MSCs and identified two subpopulations with potential functions in self-renewal and immunoregulation. Our findings advance the definition of MSCs by identifying the specific functions of their heterogeneous cellular composition, allowing for more specific and effective MSC application through the purification of their functional subpopulations.

Myelodysplastic syndromes disable support of human hematopoietic stem and progenitor cells by CD271, VCAM- 1 and CD146 expressing marrow niche cells

“…Since CD106 (VCAM-1) is an adhesion molecule that facilitates interactions with vascular endothelium, we combined these markers with VCAM-1 given the perivascular location of HSC niche cells in murine models, and because previous work showed that enrichment with VCAM-1 within CD271+ BM-MSCs identifies self-renewing cells 21 . Importantly, single cell analysis of CD271+ MSCs has identified similar functional subsets within the MSC population as those we have identified here [38][39][40][41] .…”

Fibroblasts as Turned Agents in Cancer Progression

“…Population and single-cell mRNA sequencing have demonstrated that both inter- and intra-tissue heterogeneity, gene expression, signaling pathway activation, and differentiation potentials of MSCs are exceptionally diverse, and serve the needs of the differentiated organs [ 12 , 47 , 48 ]. Single-cell transcriptomic sequencing of bone marrow and umbilical cord Wharton’s jelly MSCs revealed five distinct subpopulations that differentiate from stem-like active proliferative cells (APCs) [ 49 ]. These subpopulations express the perivascular mesodermal progenitor markers chondroitin sulfate proteoglycan 4 (CSPG4)/melanoma cell adhesion molecule-MUC18 (MCAM)/nestin (NES) of multipotent progenitor cells [ 49 ].…”

“…Single-cell transcriptomic sequencing of bone marrow and umbilical cord Wharton’s jelly MSCs revealed five distinct subpopulations that differentiate from stem-like active proliferative cells (APCs) [ 49 ]. These subpopulations express the perivascular mesodermal progenitor markers chondroitin sulfate proteoglycan 4 (CSPG4)/melanoma cell adhesion molecule-MUC18 (MCAM)/nestin (NES) of multipotent progenitor cells [ 49 ]. Subsequently, they branch off into either adipogenesis or osteochondrogenesis and differentiate into unipotent prechondrocytes specifically expressing immunomodulatory genes able to suppress activated CD3 + T cell proliferation [ 49 ].…”

Short heat shock has a long-term effect on mesenchymal stem cells’ transcriptome

“…Studies using single-cell RNA-seq have shown that MSCs are a heterogeneous population containing pre-MSCs and lineage committed MSCs [35][36][37]. Hence, the effect of environmental factors, including heat, on MSCs transcriptome and function is not yet fully understood.…”