Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage Specification
Heparan sulfate proteoglycans (HSPGs) within the neuronal niche are expressed during brain development, contributing to multiple aspects of neurogenesis, yet their roles in glial lineage commitment remain elusive. This study utilised three human cell models expanded under basal culture conditions fo...
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2025-07-01
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| author | Chieh Yu Duy L. B. Nguyen Martina Gyimesi Ian W. Peall Son H. Pham Lyn R. Griffiths Rachel K. Okolicsanyi Larisa M. Haupt |
| author_facet | Chieh Yu Duy L. B. Nguyen Martina Gyimesi Ian W. Peall Son H. Pham Lyn R. Griffiths Rachel K. Okolicsanyi Larisa M. Haupt |
| author_sort | Chieh Yu |
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| description | Heparan sulfate proteoglycans (HSPGs) within the neuronal niche are expressed during brain development, contributing to multiple aspects of neurogenesis, yet their roles in glial lineage commitment remain elusive. This study utilised three human cell models expanded under basal culture conditions followed by media-induced lineage induction to identify a reproducible and robust model of gliogenesis. SH-SY5Y human neuroblastoma cells (neuronal control), ReNcell CX human neural progenitor cells (astrocyte inductive) and ReNcell VM human neural progenitor (mixed neural induction) models were examined. The cultures were characterised during basal and inductive states via Q-PCR, Western Blotting, immunocytochemistry (ICC) and calcium signalling activity analyses. While the ReNcell lines did not produce fully mature or homogeneous astrocyte cultures, the ReNcell CX cultures most closely resembled an astrocytic phenotype with ReNcell VM cells treated with platelet-derived growth factor (PDGF) biased toward an oligodendrocyte lineage. The glycated variant of surface-bound glypican-2 (GPC2) was found to be associated with lineage commitment, with GPC6 and 6-<i>O</i> HS sulfation upregulated in astrocyte lineage cultures. Syndecan-3 (SDC3) emerged as a lineage-sensitive proteoglycan, with its cytoplasmic domain enriched in progenitor-like states and lost upon differentiation, supporting a role in maintaining neural plasticity. Conversely, the persistence of transmembrane-bound SDC3 in astrocyte cultures suggest continued involvement in extracellular signalling and proteoglycan secretion, demonstrated by increased membrane-bound HS aggregates. This data supports HSPGs and HS GAGs as human neural lineage differentiation and specification markers that may enable better isolation of human neural lineage-specific cell populations and improve our understanding of human neurogenesis. |
| format | Article |
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| institution | DOAJ |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-07-01 |
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| spelling | doaj-art-a3450d46f14647e185fd075522c4ddda2025-08-20T03:02:48ZengMDPI AGCells2073-44092025-07-011415115810.3390/cells14151158Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage SpecificationChieh Yu0Duy L. B. Nguyen1Martina Gyimesi2Ian W. Peall3Son H. Pham4Lyn R. Griffiths5Rachel K. Okolicsanyi6Larisa M. Haupt7Centre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaCentre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaCentre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaCentre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaCentre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaCentre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaCentre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaCentre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, AustraliaHeparan sulfate proteoglycans (HSPGs) within the neuronal niche are expressed during brain development, contributing to multiple aspects of neurogenesis, yet their roles in glial lineage commitment remain elusive. This study utilised three human cell models expanded under basal culture conditions followed by media-induced lineage induction to identify a reproducible and robust model of gliogenesis. SH-SY5Y human neuroblastoma cells (neuronal control), ReNcell CX human neural progenitor cells (astrocyte inductive) and ReNcell VM human neural progenitor (mixed neural induction) models were examined. The cultures were characterised during basal and inductive states via Q-PCR, Western Blotting, immunocytochemistry (ICC) and calcium signalling activity analyses. While the ReNcell lines did not produce fully mature or homogeneous astrocyte cultures, the ReNcell CX cultures most closely resembled an astrocytic phenotype with ReNcell VM cells treated with platelet-derived growth factor (PDGF) biased toward an oligodendrocyte lineage. The glycated variant of surface-bound glypican-2 (GPC2) was found to be associated with lineage commitment, with GPC6 and 6-<i>O</i> HS sulfation upregulated in astrocyte lineage cultures. Syndecan-3 (SDC3) emerged as a lineage-sensitive proteoglycan, with its cytoplasmic domain enriched in progenitor-like states and lost upon differentiation, supporting a role in maintaining neural plasticity. Conversely, the persistence of transmembrane-bound SDC3 in astrocyte cultures suggest continued involvement in extracellular signalling and proteoglycan secretion, demonstrated by increased membrane-bound HS aggregates. This data supports HSPGs and HS GAGs as human neural lineage differentiation and specification markers that may enable better isolation of human neural lineage-specific cell populations and improve our understanding of human neurogenesis.https://www.mdpi.com/2073-4409/14/15/1158heparan sulfate proteoglycansneural cell lineshuman neurogenesissyndecanglypicanlineage differentiation |
| spellingShingle | Chieh Yu Duy L. B. Nguyen Martina Gyimesi Ian W. Peall Son H. Pham Lyn R. Griffiths Rachel K. Okolicsanyi Larisa M. Haupt Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage Specification Cells heparan sulfate proteoglycans neural cell lines human neurogenesis syndecan glypican lineage differentiation |
| title | Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage Specification |
| title_full | Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage Specification |
| title_fullStr | Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage Specification |
| title_full_unstemmed | Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage Specification |
| title_short | Heparan Sulfate Proteoglycans as Potential Markers for In Vitro Human Neural Lineage Specification |
| title_sort | heparan sulfate proteoglycans as potential markers for in vitro human neural lineage specification |
| topic | heparan sulfate proteoglycans neural cell lines human neurogenesis syndecan glypican lineage differentiation |
| url | https://www.mdpi.com/2073-4409/14/15/1158 |
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