(436f) 3D Printed Microfluidic Platforms for Brain Organoid Culture

Human brain organoids are 3D millimeter and sub-millimeter scale spheroidal structures created by directing the differentiation of stem cells into neuronal and glial, which then self-organize into complex structures recapitulating the functionality of a developing brain. These organoids are currently being used to study neurodevelopmental and neurodegenerative diseases, drug development, and chemical safety testing. Since they are derived from and display characteristics that resemble the human brain, brain organoids have the potential to study growth and nutrient transport, particularly during embryogenesis, with higher human relevance than traditional animal models.

While the current generation of organoids may recapitulate spatiotemporal molecular signatures, gene expression networks, and neuron phenotypes, they do not reflect the regional organization and complexity of neuronal circuitry that allow for higher-order brain function. Our current organoids ^1,2 sustain themselves on passive diffusion of media and are 300-500 microns in size. But growing them any bigger results in starvation-derived necrosis in the core of the organoid, as confirmed by many groups growing larger organoids. Since increased size will allow the development of higher neural complexity, there is an inherent need to develop a perfusable vasculature within this organoid system to enable it to grow to sizes up to and beyond 10 mm. Here, we describe 3D printed microfluidic platforms encased within a shell for cell media perfusion and waste removal. We are designing, simulating, and optimizing microfluidic transport for optimal organoid viability. By controlling flow characteristics and chemical gradients using these microfluidic networks, we can enable stable culture and the potential to engineer neural complexity in these brain organoids of broad relevance to organoid intelligence (OI) and develop more advanced brain models to investigate chemical toxicity and screen drugs.

¹ Pamies D, Barreras P, Block K, Makri G, Kumar A, Wiersma D, et al. A human brain microphysiological system derived from induced pluripotent stem cells to study neurological diseases and toxicity. ALTEX. 2017;34(3):362–76.

² Romero JC, Berlinicke C, Chow S, Duan Y, Wang Y, Chamling X, et al. Oligodendrogenesis and myelination tracing in a CRISPR/Cas9-engineered brain microphysiological system. Front Cell Neurosci 2023