Organization of the layers recapitulates early corticogenesis observed during human brain development. (D) CTIP2, a marker of the developing cortical plate, co-localizes with β-tubulin III+ neurons in a cortical plate-like region. β-tubulin III+ neurons (green) are adjacent to the ventricular zone. ![]() (C) PAX6+ apical progenitors (red, enclosed by dotted line) are localized to a ventricular zone-like region. ![]() (B) Immunohistological analysis on cryosections of cerebral organoids reveals cortical regions within the organoid labelled by the apical progenitor marker PAX6 (red) and neuronal marker β-tubulin III (green). ![]() Cerebral organoids at this stage are made up of phase-dark structures that may be surrounded by regions of thinner, more translucent structures that display layering (arrowheads). (A) A representative phase-contrast image of a whole cerebral organoid at day 40 generated using the STEMdiff™ Cerebral Organoid Kit. Beginning with an embryoid body (EB) formation step followed by expansion of neuroepithelia, organoids generated using STEMdiff™ Cerebral Organoid Kit feature cortical-like regions including the ventricular zone (PAX6+/SOX2+/Ki-67+), outer subventricular zone (Ki-67+/p-Vimentin+), intermediate zone (TBR2+), and cortical plate (CTIP2+/MAP2+/TBR1+), which layer in similar orientations as those observed in vivo.įor extended culture periods (> 40 days), the components required for maturation can be purchased as STEMdiff™ Cerebral Organoid Maturation Kit (Catalog #08571). Science, 2014) to more reliably generate cerebral organoids. These defined, serum-free cell culture media and simple, four-stage protocol are based on the formulation published by Lancaster et al. Generate self-organized, pluripotent stem cell (PSC)-derived neural organoids with a cellular composition and structural organization representative of the developing human brain. ![]()
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