Engineering functional ventral midbrain dopaminergic neurons in human organoids through WNT modulation and bioreactor culture.

Overview

1. Institut Imagine Paris - Inserm U1163, 24 boulevard du Montparnasse, 75015 Paris, France
2. Université Paris Cité, 85 boulevard Saint-Germain, 75006 Paris, France
3. Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815 USA
4. Killion Center for Neurodegeneration and Experimental Therapeutics, University of Alabama at Birmingham, 1719 6th Ave South, Birmingham, AL 35294 USA

Abstract

Human midbrain organoids (hMOs) derived from induced pluripotent stem cells provide a powerful system to model disorders involving dopamine (DA) dysfunction, including Parkinson’s disease (PD) and neuropsychiatric conditions. However, current differentiation protocols still fall short in recapitulating early specification, substantia nigra pars compacta (SNpc)-like identity, and the functional maturation of vulnerable DA neurons. Here, we established a differentiation strategy that combines tri-phasic WNT modulation with dynamic bioreactor culture to generate hMOs enriched in SNpc-like DA neurons. This approach significantly increases the yield of TH⁺/GIRK2⁺ and TH⁺/ALDH1A1⁺ DA neurons and promotes enhanced synaptic maturation, robust electrophysiological activity, and elevated DA release. Single-cell transcriptomics revealed that this strategy drives the emergence of SOX6+/GIRK2+ SNpc-like neurons, accompanied by upregulation of synaptic, metabolic, and maturation programs, alongside reduced cell stress and apoptotic signaling. Importantly, hMOs demonstrated vulnerability upon exposure to α-synuclein preformed fibrils, resulting in aggregate formation and DA neuron degeneration, supporting their use as a human model of PD-relevant pathology. Overall, this system provides a scalable and physiologically relevant approach to investigate molecular mechanisms underlying neurodegeneration and DA-related disorders.

Reproduced under the paper's license (CC BY), from the paper cited above.

Code

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Data

Datasets cited

• geo:GSE301365, at NCBI GEO; found in “Data availability”
• zenodo:20039098, at Zenodo; found in “Data availability”

Data availability

The data, code, protocols, and key lab materials used and generated in this study are listed in a Key Resources Table alongside their persistent identifiers at the following link: https://doi.org/10.5281/zenodo.20039098. Raw single-cell RNA-seq data are available in GEO under accession number GSE301365 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE301365).

Reproduced under the paper's license (CC BY), from the paper cited above.

Versions

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Version 2, 28 September 2026

• Publisher: n/a → Springer Nature

Version 1, 27 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 6 authors, 3 keywords, 14 MeSH terms, 3 funders, 85 references, 1 RRID.

Cite

This paper

Raji, H., Bertoli, F., Perez, M. J., Lam, A., Volpicelli-Daley, L., & Deleidi, M. (2026). Engineering functional ventral midbrain dopaminergic neurons in human organoids through WNT modulation and bioreactor culture. Molecular psychiatry, 31(10), 5900-5916. https://doi.org/10.1038/s41380-026-03667-4

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