维生素C（Asc）、地塞米松（Dex）和β-甘油磷酸（β-Gly）是常用于促进离体培养内成骨细胞的成骨行为的物质。根据文献报道，一些骨肉瘤细胞系对后者的增殖动力学和成骨基因转录呈现不同反应。可想而知，这些差异导致了文献中的对比数据，需要进行初步研究以确认所选择的骨肉瘤细胞系在Asc、Dex和β-Gly存在下的表型。本研究将Saos-2细胞暴露于不同组合的Asc、Dex和β-Gly中，连续培养14天，并与人类间充质干/基质细胞（MSCs）进行对比。通过分析细胞数量、细胞毒性、矿化基质沉积和细胞增殖，评估Asc、Dex和β-Gly存在下类骨细胞样行为。此外，连续14天的培养期间，每隔两天进行一次运行相关转录因子2（RUNX2）、骨钙素（OCN）、碱性磷酸酶（ALP）、磷酸酶调控内肽酶同源物X-连锁型（PHEX）、增殖标记物MKI67和细胞增殖核抗原（PCNA）的基因表达分析。结果发现，β-Gly显著降低了Saos-2细胞的增殖，而在hMSCs中观察不到变化。此外，与hMSCs不同，Saos-2细胞显示出OCN和PHEX的上调表达，提示β-Gly可能影响骨细胞分化的后期阶段。总之，需要考虑到当将β-Gly用作类骨细胞模型时，它会显著影响Saos-2关键细胞过程的事实。© 2023. Springer Nature Limited.

Ascorbic acid (Asc), dexamethasone (Dex) and β-glycerophosphate (β-Gly) are commonly used to promote osteogenic behaviour by osteoblasts in vitro. According to the literature, several osteosarcoma cells lines appear to respond differently to the latter with regards to proliferation kinetics and osteogenic gene transcription. Unsurprisingly, these differences lead to contrasting data between publications that necessitate preliminary studies to confirm the phenotype of the chosen osteosarcoma cell line in the presence of Asc, Dex and β-Gly. The present study exposed Saos-2 cells to different combinations of Asc, Dex and β-Gly for 14 days and compared the response with immortalised human mesenchymal stromal/stem cells (MSCs). Cell numbers, cytotoxicity, mineralised matrix deposition and cell proliferation were analysed to assess osteoblast-like behaviour in the presence of Asc, Dex and β-Gly. Additionally, gene expression of runt-related transcription factor 2 (RUNX2); osteocalcin (OCN); alkaline phosphatase (ALP); phosphate regulating endopeptidase homolog X-linked (PHEX); marker of proliferation MKI67 and proliferating cell nuclear antigen (PCNA) was performed every two days during the 14-day cultures. It was found that proliferation of Saos-2 cells was significantly decreased by the presence of β-Gly which contrasted with hMSCs where no change was observed. Furthermore, unlike hMSCs, Saos-2 cells demonstrated an upregulated expression of late osteoblastic markers, OCN and PHEX that suggested β-Gly could affect later stages of osteogenic differentiation. In summary, it is important to consider that β-Gly significantly affects key cell processes of Saos-2 when using it as an osteoblast-like cell model.© 2023. Springer Nature Limited.