Expression of neural cell adhesion molecule and polysialic acid in human bone marrow-derived mesenchymal stromal cells

Abstract

BackgroundIn order to develop novel clinical applications and to gain insights into possible therapeutic mechanisms, detailed molecular characterization of human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) is needed. Neural cell adhesion molecule (NCAM, CD56) is a transmembrane glycoprotein modulating cell–cell and cell–matrix interactions. An additional post-translational modification of NCAM is the α2,8-linked polysialic acid (polySia). Because of its background, NCAM is often considered a marker of neural lineage commitment. Generally, hBM-MSCs are considered to be devoid of NCAM expression, but more rigorous characterization is needed.MethodsWe have studied NCAM and polySia expression in five hBM-MSC lines at mRNA and protein levels. Cell surface localization was confirmed by immunofluorescence staining and expression frequency in the donor-specific lines by flow cytometry. For the detection of poorly immunogenic polySia, a fluorochrome-tagged catalytically defective enzyme was employed.ResultsAll five known NCAM isoforms are expressed in these cells at mRNA level and the three main isoforms are present at protein level. Both polysialyltransferases, generally responsible for NCAM polysialylation, are expressed at mRNA level, but only very few cells express polySia at the cell surface.ConclusionsOur results underline the need for a careful control of methods and conditions in the characterization of MSCs. This study shows that, against the generally held view, clinical-grade hBM-MSCs do express NCAM. In contrast, although both polysialyltransferase genes are transcribed in these cells, very few express polySia at the cell surface. NCAM and polySia represent new candidate molecules for influencing MSC interactions.

Internalization of a polysialic acid-binding Escherichia coli bacteriophage into eukaryotic neuroblastoma cells

“…In culture, the cell line contains two morphological variants, the neuroblastic (N-type) cells rich in polysialic acid as well as the flat substrate-adherent/Schwannian (S-type) cells containing little polysialic acid 30 . A polysialic acid and NCAM-expressing N-type cell population (kSK-N-SH) has been previously isolated from the SK-N-SH cell line 31 . The labeled PK1A2 phages bound specifically to the N-type cells and had a similar distribution to the polysialic acid carrier molecule NCAM (Fig.…”

“…The cell lines obtained from the American Type Culture Collection (ATCC) included human neuroblastoma cell lines SK-N-SH (ATCC HTB-11), SK-N-AS (ATCC CRL-2137) and SH-SY5Y (ATCC CRL-2266), human foreskin fibroblast cell line BJ (ATCC CRL-2522), and baby hamster kidney fibroblast cell line BHK-21 (ATCC CCL-10). The polysialic acid-expressing cell line kSK-N-SH was previously established in our laboratory from SK-N-SH cells by immunomagnetic bead separation 31 , and was authenticated together with SK-N-SH cell line by using GenePrint® 10 system (Institute for Molecular Medicine Finland). Cell lines were checked for the absence of mycoplasma infections using the EZ-PCR Mycoplasma Test Kit (Biological Industries).…”

Development, characterization and virus susceptibility of a continuous cell line from the caudal fin of marbled eel (Anguilla marmorata)

“…Nestin is an intermediate filament protein expressed in stem and progenitor cells of neuroectodermal, endodermal and mesodermal lineages (Wiese et al, 2004). Ncam1 (Cd56) is commonly considered a specific cell surface marker for neurons and a subset of mammalian NK cells, yet it also appears in epithelial and mesenchymal stem and progenitor cells including those of the epidermis, liver, kidney and muscles (Buzhor et al, 2013;Skog et al, 2016;Terada, 2013;Trapecar, Kelc, Gradisnik, Vogrin, & Rupnik, 2014;Wang et al, 2018). Cd146 (a melanoma cell adhesion molecule) is a marker of endothelial cells that is also expressed in perivascular mesenchymal stem cells (pericytes) as well as muscle stem and progenitor cells (Crisan et al, 2008;Lecourt et al, 2010).…”

Persistent development of adomavirus and aquareovirus in a novel cell line from marbled eel with petechial skin haemorrhage

“…PCR analysis demonstrated that they exhibit a gene expression pattern (i.e., only nestin and ncam1 genes) different from that of MECF‐1 cells, which express transcripts of the cd146, ictacalcin, k13, myod1, ncam1 and nestin genes (Pao et al, ). Nestin and NCAM1 are markers of epithelial and mesenchymal stem and progenitor cells (Buzhor et al, ; Skog et al, ; Wiese et al, ). The result proposes that DMEPF‐5 harbours the stem and progenitor cells; further characterization of DMEPF‐5 cells is ongoing.…”