In vitro modeling of the human dopaminergic system using spatially arranged ventral midbrain–striatum–cortex assembloids

• Published in: Nature methods Vol. 20; no. 12; pp. 2034 - 2047
• Main Authors: Reumann, Daniel, Krauditsch, Christian, Novatchkova, Maria, Sozzi, Edoardo, Wong, Sakurako Nagumo, Zabolocki, Michael, Priouret, Marthe, Doleschall, Balint, Ritzau-Reid, Kaja I., Piber, Marielle, Morassut, Ilaria, Fieseler, Charles, Fiorenzano, Alessandro, Stevens, Molly M., Zimmer, Manuel, Bardy, Cedric, Parmar, Malin, Knoblich, Jürgen A.
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01-12-2023; Nature Publishing Group
• Subjects: 631/378/1689; 631/378/2571; 631/532/1360; 692/308/1426; 692/308/2171; Addictions; Basic Medicine; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Cell therapy; Circuits; Cocaine; Cognition; Corpus Striatum; Cortex; Dopamine; Dopamine receptors; Dopaminergic Neurons; Drug abuse; Embedding; Humans; Immunosuppressive agents; Innervation; Life Sciences; Medical and Health Sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Mental disorders; Mesencephalon; Mesencephalon - anatomy & histology; Mesencephalon - physiology; Movement disorders; Narcotics; Neostriatum; Neurodegeneration; Neurodegenerative diseases; Neurons; Neurosciences; Neurovetenskaper; Organoids; Parkinson Disease; Parkinson's disease; Proteomics; Transplantation; Withdrawal
• ISSN: 1548-7091; 1548-7105; 1548-7105
• DOI: 10.1038/s41592-023-02080-x

Ventral midbrain dopaminergic neurons project to the striatum as well as the cortex and are involved in movement control and reward-related cognition. In Parkinson’s disease, nigrostriatal midbrain dopaminergic neurons degenerate and cause typical Parkinson’s disease motor-related impairments, while the dysfunction of mesocorticolimbic midbrain dopaminergic neurons is implicated in addiction and neuropsychiatric disorders. Study of the development and selective neurodegeneration of the human dopaminergic system, however, has been limited due to the lack of an appropriate model and access to human material. Here, we have developed a human in vitro model that recapitulates key aspects of dopaminergic innervation of the striatum and cortex. These spatially arranged ventral midbrain–striatum–cortical organoids (MISCOs) can be used to study dopaminergic neuron maturation, innervation and function with implications for cell therapy and addiction research. We detail protocols for growing ventral midbrain, striatal and cortical organoids and describe how they fuse in a linear manner when placed in custom embedding molds. We report the formation of functional long-range dopaminergic connections to striatal and cortical tissues in MISCOs, and show that injected, ventral midbrain-patterned progenitors can mature and innervate the tissue. Using these assembloids, we examine dopaminergic circuit perturbations and show that chronic cocaine treatment causes long-lasting morphological, functional and transcriptional changes that persist upon drug withdrawal. Thus, our method opens new avenues to investigate human dopaminergic cell transplantation and circuitry reconstruction as well as the effect of drugs on the human dopaminergic system.
MISCOs are spatially arranged ventral midbrain–striatum–cortical organoids that enable investigations into the human dopaminergic system.