Suspension culture drives metabolic rewiring in human pluripotent stem cells

Transitioning human pluripotent stem cells from 2D to 3D suspension culture rewires metabolism, with 3D showing increased glucose-derived pyruvate labeling and a higher lactate/glucose ratio.

• Why it matters: Scalable 3D suspension expansion is common, but the molecular and metabolic consequences of moving from 2D remain unclear.
• What they did: The authors compared 2D versus 3D hPSC cultures using transcriptomics, proteomics, spent-medium profiling, and stable-isotope tracing to track glucose carbon flow.
• The result: Multi-omics identified culture format as the dominant molecular driver and supported increased glycolysis in 3D, alongside reduced glutamine contribution to the TCA cycle and increased relative reductive carboxylation.

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