Abstract
Brain organoids derived from human pluripotent stem cells provide a highly valuable in vitro model to recapitulate human brain development and neurological diseases. However, the current systems for brain organoid culture require further improvement for the reliable production of high-quality organoids. Here, we demonstrate two engineering elements to improve human brain organoid culture, (1) a human brain extracellular matrix to provide brain-specific cues and (2) a microfluidic device with periodic flow to improve the survival and reduce the variability of organoids. A three-dimensional culture modified with brain extracellular matrix significantly enhanced neurogenesis in developing brain organoids from human induced pluripotent stem cells. Cortical layer development, volumetric augmentation, and electrophysiological function of human brain organoids were further improved in a reproducible manner by dynamic culture in microfluidic chamber devices. Our engineering concept of reconstituting brain-mimetic microenvironments facilitates the development of a reliable culture platform for brain organoids, enabling effective modeling and drug development for human brain diseases.
Highlights
Brain organoids derived from human pluripotent stem cells provide a highly valuable in vitro model to recapitulate human brain development and neurological diseases
Our bioengineered human cerebral organoids exhibit structural, phenotypic, and functional features being observed during whole human brain development as follows; (1) the structural features of spontaneous brain morphogenesis, (2) mature neuronal identities (e.g. abundant expression of mature neuronal markers (NeuN, vesicular glutamate transporter 1 (VGLUT1), GAD, GABA), extensive neural networks (Ncadherin), and synaptogenesis (SYNI, PSD95) at earlier time points), (3) divergent brain cell population, and (4) electrophysiologically active properties
A recent study reported that the expression of mature neuronal markers, including VGLUT1 and Glutamate decarboxylase 1 (GAD1), was observed in 6-month-old cerebral organoids[69]
Summary
Brain organoids derived from human pluripotent stem cells provide a highly valuable in vitro model to recapitulate human brain development and neurological diseases. Due to the lack of instructive signals during the generation of human cerebral organoids, they recapitulate only some of the earliest stages of human embryonic brain development[5] and are not able to mimic the later stages of neurogenesis until extended cultivation for 6–9 months[19] Another critical limitation is the extensive cell death in the developing organoids at later stages due to diffusional limitations in oxygen and nutrient transfer[4,8]. We reason that providing brain-specific extracellular matrix (ECM) cues together with improved nutrient and oxygen exchange will support cell expansion as well as neuronal differentiation and functional maturation, thereby recapitulating prominent features of human embryonic cortical development in a much precise and reproducible manner
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