Cell-Surface Expression of Neuron-Glial Antigen 2 (NG2) and Melanoma Cell Adhesion Molecule (CD146) in Heterogeneous Cultures of Marrow-Derived Mesenchymal Stem Cells

Abstract

Cellular heterogeneity of mesenchymal stem cells (MSCs) impedes their use in regenerative medicine. The objective of this research is to identify potential biomarkers for the enrichment of progenitors from heterogeneous MSC cultures. To this end, the present study examines variation in expression of neuron-glial antigen 2 (NG2) and melanoma cell adhesion molecule (CD146) on the surface of MSCs derived from human bone marrow in response to culture conditions and among cell populations. Multipotent cells isolated from heterogeneous MSC cultures exhibit a greater than three-fold increase in surface expression for NG2 and greater than two-fold increase for CD146 as compared with parental and lineage-committed MSCs. For both antigens, surface expression is downregulated by greater than or equal to six-fold when MSCs become confluent. During serial passage, maximum surface expression of NG2 and CD146 is associated with minimum doubling time. Upregulation of NG2 and CD146 during loss of adipogenic potential at early passage suggests some limits to their utility as potency markers. A potential relationship between proliferation and antigen expression was explored by sorting heterogeneous MSCs into rapidly and slowly dividing groups. Fluorescence-activated cell sorting revealed that rapidly dividing MSCs display lower scatter and 50% higher NG2 surface expression than slowly dividing cells, but CD146 expression is comparable in both groups. Heterogeneous MSCs were sorted based on scatter properties and surface expression of NG2 and CD146 into high (HI) and low (LO) groups. Sc LO NG2 HI and Sc LO NG2 HI CD146 HI MSCs have the highest proliferative potential of the sorted groups, with colony-forming efficiencies that are 1.5-2.2 times the value for the parental controls. The Sc LO gate enriches for rapidly dividing cells. Addition of the NG2 HI gate increases cell survival to 1.5 times the parental control. Further addition of the CD146 HI gate does not significantly improve cell division or survival. The combination of low scatter and high NG2 surface expression is a promising selection criterion to enrich a proliferative phenotype from heterogeneous MSCs during ex vivo expansion, with potentially numerous applications.

Nestin+ kidney resident mesenchymal stem cells for the treatment of acute kidney ischemia injury

“…In this study, renal Nestin þ cells expressed some mesenchymal markers, such as Sca-1, CD44, CD106, but lack of expression CD45 and CD11b by FACS analysis. By immunofluorescent staining, these cells co-expressed Nestin with vimentin [42], NG2 [43] and PDGFRa [27], which were all reported as potential markers of MSCs.…”

NOTCH3 regulates stem-to–mural cell differentiation in infantile hemangioma

“…Although PDGFRβ and NG2 are considered mural cell markers in the vasculature (21,22), both are expressed in progenitor cells. NG2 expression has been reported in bone marrow mesenchymal stem cells (29,30), neuronal progenitor cells (31,32), and lymphatic malformation progenitor cells (33). Similarly, PDGFRβ is expressed in circulating endothelial progenitors, as well as hemangioblasts (34,35), suggesting that HemECs have failed to terminally differentiate from endothelial progenitor cells.…”

Receptors and Binding Structures for Clostridium difficile Toxins A and B

“…CSPG4/NG2 is expressed in tissues of brain, gastrointestinal tract, endocrine organs, and >50 cell types (Levine and Nishiyama 1996;Nicolosi et al 2015). This cell surface protein is characteristic for progenitor cells of epithelial and mesodermal origin (Russell et al 2013). The ectodomain of CSPG4 binds to type V and VI collagens, multi-PDZ domain proteins, and others (Stallcup et al 1990;Nishiyama et al 1996;Tillet et al 1997).…”