Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells

• DOI: 10.3389/fcell.2024.1227034
• Publication Date: 2024-02-22T04:23:39Z
• Authors: Agnieszka Smolinska; Magdalena Chodkowska; Agata Kominek; Jakub Janiec; Katarzyna Piwocka; Dorota Sulejczak; Anna Sarnowska
• Keywords: Cell Sorting; Wharton's jelly; Stem cell marker; Lineage markers

Background: High heterogeneity of mesenchymal stem/stromal cells (MSCs) due to different degrees differentiation cell subpopulations poses a considerable challenge in preclinical studies. The at pluripotent-like stage represent stem population interest for many researchers worldwide, which is worthy identification, isolation, and functional characterization. In the current study, we asked whether Wharton’s jelly-derived MSCs (WJ-MSCs) express stage-specific embryonic antigen-4 (SSEA-4) can be considered as population. Methods: SSEA-4 expression culture conditions was compared efficiency two separation methods were assessed: Magnetic Activated Cell Sorting (MACS) Fluorescence (FACS). After SSEA-4+ analyzed following parameters: maintenance antigen after sorting, cell-related gene expression, proliferation potential, clonogenicity, secretome profiling, ability form spheres under 3D conditions. Results: FACS allowed enrichment content that lasted six passages sorting. Despite elevated stemness-related genes, neither differed their clonogenicity potential from initial negative populations nor exhibited pluripotent repertoire. observed smaller spheroids increased survival Conclusion: transient our findings could not fully confirm undifferentiated nature WJ-MSC cultured vitro .