Neural organoids are invaluable model systems for studying neurodevelopment, generated by either guided or unguided approaches. Despite the importance for the field, the resulting differences between these models are unclear. To obtain an unbiased comparison, we performed a multi-omic analysis of forebrain organoids generated in parallel with two widely applied guided and unguided protocols. The guided forebrain organoids contained a larger proportion of neurons, including GABAergic interneurons, whereas the unguided organoids contained significantly more choroid plexus, radial glia, and astrocytes at later stages. Substantial differences in metabolic profiles were identified, pointing to increased levels of oxidative phosphorylation and fatty acid β-oxidation in the unguided forebrain organoids and a higher reliance on glycolysis in the guided forebrain organoids. Overall, our study comprises a thorough description of the multi-omic differences between these guided and unguided forebrain organoids and provides an important resource for the neural organoid field studying neurodevelopment and disease.
Øhlenschlæger et al. (2026) studied this question.