Cerebral organoids, three-dimensional cultures that model organogenesis, provide a new platform

Cerebral organoids, three-dimensional cultures that model organogenesis, provide a new platform to investigate human brain development. cell type in the body to model human development and disease, screen for therapeutic drugs, and develop cell replacement therapies. Traditional monolayer cultures allow for external control of targeted differentiation of human iPSCs to produce more uniform cell populations; however, these cultures lack 3D cell assembly properties that define endogenous biological systems. Structures resembling whole developing organs, named organoids, have recently been generated via 3D cultures and include intestinal, kidney, retinal, and cerebral organoids (Lancaster and Knoblich, 2014; Yin et al., 2016). Organoid technology developed from embryoid body cultures, which are 3D aggregates of stem cells that self-organize to develop disparate tissues in vitro, comparable to teratoma formation in vivo. Organoids provide a unique opportunity to model human organogenesis, which is usually not accessible to experimentation. An immediate application of organoid technology would be to address the current global public health emergency concerning a suspected link between Zika computer virus (ZIKV) and microcephaly, a neurodevelopmental disorder, by modeling human brain development. One recent advance in cerebral organoid technology was the ownership of a spinning bioreactor to facilitate nutrient and oxygen absorption, which enables formation of longer neuroepithelium-like zones and supports growth of large, complex organoids that more closely resemble Rabbit Polyclonal to MRPL14 the developing human brain than experienced been achieved by previous methods (Lancaster et al., 2013). Derived from an early NASA-designed rotating wall ship bioreactor to simulate microgravity, this technology potentially offers two additional benefits: low fluid shear stress to promote cell-cell interactions and induction of differentiation (Goodwin et al., 1993); and randomized gravitational vectors that impact intracellular transmission transduction and gene manifestation (Jessup et al., 1993). This and other human cerebral organoid technologies (Kadoshima et al., 2013; Lancaster et al., 2013; Mariani et al., 2015; Muguruma et al., 2015; Pasca 184475-55-6 IC50 et al., 2015) have generated much enjoyment for using organoids to model human brain development and disorders. Despite the 184475-55-6 IC50 promise of these pioneering organoid technologies, there are several major difficulties. First, available spinning bioreactors require a large volume of medium and incubator space (Physique H1Aa). With frequent media changes over several months of culturing, the system is usually cost-prohibitive for most laboratories and precludes scalability, use of growth factors, or chemical screening. It also presents a roadblock for screening different conditions to optimize protocols. Second, the current cerebral organoid strategy (intrinsic protocol) is usually based on cell self-assembly without external control, and thus each organoid is usually typically comprised of diverse cell types found in forebrain, hindbrain, and retina (Lancaster et al., 2013). Large sample to sample variability associated with current methods complicates quantitative analyses and limits applicability. Third, important features of human brain development have yet to be robustly recapitulated in cerebral organoids. For example, unlike rodents, the embryonic human cerebral cortex contains an abundant populace of specialized outer radial glia cells (oRGCs) in the outer subventricular zone (oSVZ), the cellular populace considered pivotal to the 184475-55-6 IC50 evolutionary increase in human cortex size and complexity (Lui et al., 2011). Current cerebral organoids contain only sparse progenitors that have morphological characteristics of oRGCs 184475-55-6 IC50 and none have exhibited a well-developed oSVZ layer. Taken together, there is usually a crucial need to develop an organoid platform with reduced cost, higher throughput, increased reproducibility, and that better resembles crucial aspects of human cortical development. To address these challenges, we designed a miniaturized spinning bioreactor using 3D design and printing technology, and developed a protocol to generate forebrain-specific organoids from human iPSCs, which recapitulate human embryonic cortical development in a reproducible and quantifiable manner. We also developed protocols for midbrain and hypothalamic organoids. For proof-of-principle applications of our platform, 184475-55-6 IC50 we.

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