Microfluidic device with brain extracellular matrix promotes structural and functional maturation of human brain organoids

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.

Bio-inspired engineering of a perfusion culture platform for guided three-dimensional nerve cell growth and differentiation

“…Three representative perfusion culture platforms have been developed to realize perfusion 3D cell culture, including a membrane-based multilayer device, 25 a device based on hydrogels with a built-in vascular network, 26 and a porous chamber device. 27 However, because of the limited assembly accuracy and structure complexity of the hydrogels in these perfusion platforms, the directional cues to guide oriented cell growth are difficult to generate. A two-photon lithography (TPL) technique provides an alternative approach for the one-stop fabrication of a tailor-made cell culture platform with sub-micron precision.…”

The Role of the Extracellular Matrix in Neural Progenitor Cell Proliferation and Cortical Folding During Human Neocortex Development

“…Human iPSCs grown on ECM secreted by NPCs increased the expression of neuronal genes, such as tubulin beta 3, in comparison to ECM secreted by ESCs (Yan et al, 2015 ). ECM extracted from adult human brain tissue promoted increased laminin deposition in cerebral organoids, increased proliferation of APs and increased neural production (Cho et al, 2021 ). It would be interesting to investigate the effect of ECM extracted from fetal neocortex tissue using this model, as it is highly likely to differ in composition to the adult ECM extract used in this study.…”

3D hydrogel models of the neurovascular unit to investigate blood–brain barrier dysfunction

“…Vascularisation has also been achieved by using 3D-printing of a microfluidic chip to ensure precise spatial locations, and thereby interactions, between neural organoids and vascular tissue [ 204 ]. Combining a microfluidic device with a decellularised human brain tissue-derived ECM was used to recreate brain-mimetic niches necessary to guide neural and glial differentiation for brain organogenesis [ 205 ]. These examples clearly indicate that despite the current limitations of organoid models, by combining them with hydrogel, microfluidic and 3D-bioprinting technologies these models are clearly bringing us closer to being able to replicate the complexity of the NVU in vitro .…”