Yu Chieh, Nguyen Duy L B, Gyimesi Martina, Peall Ian W, Pham Son H, Griffiths Lyn R, Okolicsanyi Rachel K, Haupt Larisa M
Centre for Genomics & Personalised Health, Genomics Research Centre, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, Australia.
ARC Training Centre for Cell and Tissue Engineering Technologies, Queensland University of Technology (QUT), Kelvin Grove, Brisbane, QLD 4059, Australia.
Cells. 2025 Jul 26;14(15):1158. doi: 10.3390/cells14151158.
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- 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.
神经小生境中的硫酸乙酰肝素蛋白聚糖（HSPGs）在大脑发育过程中表达，对神经发生的多个方面有贡献，但其在胶质细胞谱系定向中的作用仍不清楚。本研究利用三种在基础培养条件下扩增的人类细胞模型，随后通过培养基诱导谱系诱导，以确定一种可重复且强大的胶质细胞生成模型。研究了SH-SY5Y人神经母细胞瘤细胞（神经元对照）、ReNcell CX人神经祖细胞（星形胶质细胞诱导）和ReNcell VM人神经祖细胞（混合神经诱导）模型。通过定量聚合酶链反应（Q-PCR）、蛋白质免疫印迹、免疫细胞化学（ICC）和钙信号活性分析，对基础状态和诱导状态下的培养物进行了表征。虽然ReNcell系未产生完全成熟或同质的星形胶质细胞培养物，但ReNcell CX培养物最接近星形胶质细胞表型，用血小板衍生生长因子（PDGF）处理的ReNcell VM细胞偏向少突胶质细胞谱系。发现表面结合的磷脂酰肌醇蛋白聚糖-2（GPC2）的糖化变体与谱系定向有关，在星形胶质细胞谱系培养物中，GPC6和6-硫酸乙酰肝素上调。Syndecan-3（SDC3）是一种谱系敏感的蛋白聚糖，其胞质结构域在祖细胞样状态下富集，分化时丢失，支持其在维持神经可塑性中的作用。相反，跨膜结合的SDC3在星形胶质细胞培养物中的持续存在表明其继续参与细胞外信号传导和蛋白聚糖分泌，膜结合的硫酸乙酰肝素聚集体增加证明了这一点。这些数据支持HSPGs和硫酸乙酰肝素糖胺聚糖（HS GAGs）作为人类神经谱系分化和特化标志物，这可能有助于更好地分离人类神经谱系特异性细胞群体，并增进我们对人类神经发生的理解。
