# Perfusable brain organoids: a long-term culture approach

> **NIH NIH R21** · YALE UNIVERSITY · 2020 · $209,375

## Abstract

Neuropsychiatric disorders have been primarily studied from post-mortem brain samples, given the inability to
isolate neurons from living affected individuals. Unfortunately post-mortem samples offer a limited view of the
disorders, for the disorder onset is typically several decades earlier than the brain sample. In addition, cellular
physiology is difficult to analyze in postmortem tissue, limiting the experimental design. As a result, progress
toward understanding the cellular and molecular mechanisms behind neuropsychiatric disorders has been poor.
We will take advantage of recent discoveries, which have demonstrated that iPSC derived 3D cerebral
organoid culture allows large variety of neuronal specification mimicking human brain development. These
efforts have been greatly limited by necrosis near the center of the 3D spheroids due to poor metabolite
exchange with the inner most cells, and overall limited cell culture survival.
We propose to transplant the organoids in mouse as a way to achieve long-term culture conditions, via active
blood perfusion through the host vascular system. We expect the host vascular system to vascularize the
transplanted organoids. We propose to reduce the time to vascularization by developing a vasculature in the
organoids using human endothelial cells and mesenchymal stem cells. This has the added benefit of limiting
the number of mouse cells in the human organoid. We optimize culture condition pre-implantation by culturing
the organoids in a bioreactor system. We will also compare long-term culture in the bioreactor, where culture
conditions are optimal, with the transplanted organoids. The expectation is that vascular co-culture and
transplantation in mouse will generate more mature and viable neural tissue the bioreactor culture. Finally, we
will characterize the iPSC-derived 3D neural tissue by 3D immunostaining, individual organoid bulk and single
cell RNAseq.
The impact of these combined experiments will be the definition of a simple and medium to high throughput
methodology for long-term neuronal differentiation assays, aimed at producing mature neurons of diverse sub-
populations and a higher degree of structural differentiation mimicking human telencephalic development, with
potential implications for neurodevelopmental disorders, like Autism, and potentially also disorders like
Schizophrenia, Alzheimer and Parkinson disease and related drug discovery.

## Key facts

- **NIH application ID:** 9973237
- **Project number:** 5R21MH119503-02
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Gianfilippo Coppola
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $209,375
- **Award type:** 5
- **Project period:** 2019-07-05 → 2022-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9973237

## Citation

> US National Institutes of Health, RePORTER application 9973237, Perfusable brain organoids: a long-term culture approach (5R21MH119503-02). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/9973237. Licensed CC0.

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