Collection Citation: Hariam Raji, michela.deleidi 2025. Engineering functional ventral midbrain dopaminergic neurons in human organoids through WNT modulation and bioreactor culture. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlkyj7xg5r/v1
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection of protocols for midbrain organoid generation, handling and analysis.
Created: November 17, 2025
Last Modified: November 17, 2025
Collection Integer ID: 232701
Keywords: generation of human midbrain organoid, human midbrain organoid, treatment of midbrain organoid, midbrain organoid, neuronal activity in organoid, functional ventral midbrain dopaminergic neuron, apoptosis in organoid section, organoid activity, human organoid, mea recording of organoid activity, term exposure of organoid, confocal imaging of organoid tissue, organoid, organoid tissue, organoid section, immunohistochemistry, midbrain, apoptosi, confocal imaging
Abstract
This protocol collection includes the following protocols:
Human iPSC Culture: Describes feeder-free maintenance and passaging of human iPSCs to preserve pluripotency for downstream differentiation.
Generation of Human Midbrain Organoids: Outlines stepwise differentiation of iPSCs into midbrain organoids using defined patterning cues and dynamic culture.
Immunohistochemistry and Imaging: Provides a workflow for fixation, sectioning, antibody staining, and confocal imaging of organoid tissue.
Fluorometric TUNEL Assay: Details fluorescent TUNEL labeling and imaging to quantify apoptosis in organoid sections.
Treatment of Midbrain Organoids with αSyn-PFFs: Describes controlled short-term exposure of organoids to αSyn-PFFs with matched vehicle controls.
MEA Recording of Organoid Activity: Provides a procedure for measuring spontaneous and drug-modulated neuronal activity in organoids using MEA systems.
