Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells
Background: High heterogeneity of mesenchymal stem/stromal cells (MSCs) due to different degrees of differentiation of cell subpopulations poses a considerable challenge in preclinical studies. The cells at a pluripotent-like stage represent a stem cell population of interest for many researchers wo...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-02-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2024.1227034/full |
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| author | Agnieszka Smolinska Magdalena Chodkowska Agata Kominek Jakub Janiec Katarzyna Piwocka Dorota Sulejczak Anna Sarnowska |
| author_facet | Agnieszka Smolinska Magdalena Chodkowska Agata Kominek Jakub Janiec Katarzyna Piwocka Dorota Sulejczak Anna Sarnowska |
| author_sort | Agnieszka Smolinska |
| collection | DOAJ |
| description | Background: High heterogeneity of mesenchymal stem/stromal cells (MSCs) due to different degrees of differentiation of cell subpopulations poses a considerable challenge in preclinical studies. The cells at a pluripotent-like stage represent a stem cell population of interest for many researchers worldwide, which is worthy of identification, isolation, and functional characterization. In the current study, we asked whether Wharton’s jelly-derived MSCs (WJ-MSCs) which express stage-specific embryonic antigen-4 (SSEA-4) can be considered as a pluripotent-like stem cell population.Methods: SSEA-4 expression in different culture conditions was compared and the efficiency of two cell separation methods were assessed: Magnetic Activated Cell Sorting (MACS) and Fluorescence Activated Cell Sorting (FACS). After isolation, SSEA-4+ cells were analyzed for the following parameters: the maintenance of the SSEA-4 antigen expression after cell sorting, stem cell-related gene expression, proliferation potential, clonogenicity, secretome profiling, and the ability to form spheres under 3D culture conditions.Results: FACS allowed for the enrichment of SSEA-4+ cell content in the population that lasted for six passages after sorting. Despite the elevated expression of stemness-related genes, SSEA-4+ cells neither differed in their proliferation and clonogenicity potential from initial and negative populations nor exhibited pluripotent differentiation repertoire. SSEA-4+ cells were observed to form smaller spheroids and exhibited increased survival under 3D conditions.Conclusion: Despite the transient expression of stemness-related genes, our findings could not fully confirm the undifferentiated pluripotent-like nature of the SSEA-4+ WJ-MSC population cultured in vitro. |
| first_indexed | 2024-03-07T23:06:58Z |
| format | Article |
| id | doaj.art-533db5505d914317be081a18f70a44e9 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-03-07T23:06:58Z |
| publishDate | 2024-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-533db5505d914317be081a18f70a44e92024-02-22T04:23:26ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2024-02-011210.3389/fcell.2024.12270341227034Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cellsAgnieszka Smolinska0Magdalena Chodkowska1Agata Kominek2Jakub Janiec3Katarzyna Piwocka4Dorota Sulejczak5Anna Sarnowska6Translational Platform for Regenerative Medicine, Mossakowski Medical Research Institute, Polish Academy of Sciences, Warsaw, PolandTranslational Platform for Regenerative Medicine, Mossakowski Medical Research Institute, Polish Academy of Sciences, Warsaw, PolandLaboratory of Cytometry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, PolandLaboratory of Cytometry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, PolandLaboratory of Cytometry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, PolandDepartment of Experimental Pharmacology, Mossakowski Medical Research Institute, Polish Academy of Sciences, Warsaw, PolandTranslational Platform for Regenerative Medicine, Mossakowski Medical Research Institute, Polish Academy of Sciences, Warsaw, PolandBackground: High heterogeneity of mesenchymal stem/stromal cells (MSCs) due to different degrees of differentiation of cell subpopulations poses a considerable challenge in preclinical studies. The cells at a pluripotent-like stage represent a stem cell population of interest for many researchers worldwide, which is worthy of identification, isolation, and functional characterization. In the current study, we asked whether Wharton’s jelly-derived MSCs (WJ-MSCs) which express stage-specific embryonic antigen-4 (SSEA-4) can be considered as a pluripotent-like stem cell population.Methods: SSEA-4 expression in different culture conditions was compared and the efficiency of two cell separation methods were assessed: Magnetic Activated Cell Sorting (MACS) and Fluorescence Activated Cell Sorting (FACS). After isolation, SSEA-4+ cells were analyzed for the following parameters: the maintenance of the SSEA-4 antigen expression after cell sorting, stem cell-related gene expression, proliferation potential, clonogenicity, secretome profiling, and the ability to form spheres under 3D culture conditions.Results: FACS allowed for the enrichment of SSEA-4+ cell content in the population that lasted for six passages after sorting. Despite the elevated expression of stemness-related genes, SSEA-4+ cells neither differed in their proliferation and clonogenicity potential from initial and negative populations nor exhibited pluripotent differentiation repertoire. SSEA-4+ cells were observed to form smaller spheroids and exhibited increased survival under 3D conditions.Conclusion: Despite the transient expression of stemness-related genes, our findings could not fully confirm the undifferentiated pluripotent-like nature of the SSEA-4+ WJ-MSC population cultured in vitro.https://www.frontiersin.org/articles/10.3389/fcell.2024.1227034/fullmesenchymal stem/stromal cellsSSEA-4stemnesspluripotentFACSMACs |
| spellingShingle | Agnieszka Smolinska Magdalena Chodkowska Agata Kominek Jakub Janiec Katarzyna Piwocka Dorota Sulejczak Anna Sarnowska Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells Frontiers in Cell and Developmental Biology mesenchymal stem/stromal cells SSEA-4 stemness pluripotent FACS MACs |
| title | Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells |
| title_full | Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells |
| title_fullStr | Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells |
| title_full_unstemmed | Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells |
| title_short | Stemness properties of SSEA-4+ subpopulation isolated from heterogenous Wharton’s jelly mesenchymal stem/stromal cells |
| title_sort | stemness properties of ssea 4 subpopulation isolated from heterogenous wharton s jelly mesenchymal stem stromal cells |
| topic | mesenchymal stem/stromal cells SSEA-4 stemness pluripotent FACS MACs |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2024.1227034/full |
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