STEMdiff™ Cerebral Organoid Kit

Culture medium kit for establishment and maturation of human cerebral organoids

STEMdiff™ Cerebral Organoid Kit

Culture medium kit for establishment and maturation of human cerebral organoids

STEMdiff™ Cerebral Organoid Maturation Kit
1 Kit
134 USD
Catalog # 08571

Culture medium kit for extended maturation of human cerebral organoids

STEMdiff™ Cerebral Organoid Kit
1 Kit
391 USD
Catalog # 08570

Culture medium kit for establishment and maturation of human cerebral organoids

What's Included

  • STEMdiff™ Cerebral Organoid Kit (Catalog #08570)
    • STEMdiff™ Cerebral Organoid Basal Medium 1,100 mL
    • STEMdiff™ Cerebral Organoid Basal Medium 2, 250 mL
    • STEMdiff™ Cerebral Organoid Supplement A, 10 mL
    • STEMdiff™ Cerebral Organoid Supplement B, 0.5 mL
    • STEMdiff™ Cerebral Organoid Supplement C, 0.25 mL
    • STEMdiff™ Cerebral Organoid Supplement D, 0.5 mL
    • STEMdiff™ Cerebral Organoid Supplement E, 4.5 mL
  • STEMdiff™ Cerebral Organoid Maturation Kit (Catalog #08571)
    • STEMdiff™ Cerebral Organoid Basal Medium 2, 250 mL
    • STEMdiff™ Cerebral Organoid Supplement E, 4.5 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Product Information Sheet.

Overview

Generate self-organized, pluripotent stem cell (PSC)-derived neural organoids with a cellular composition and structural organization representative of the developing human brain.

These defined, serum-free cell culture media and simple, four-stage protocol are based on the formulation published by Lancaster et al. (Lancaster MA et al. Nature, 2013 and Lancaster MA et al. Science, 2014) to more reliably generate cerebral organoids. 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.

For extended culture periods (> 40 days), the components required for maturation can be purchased as STEMdiff™ Cerebral Organoid Maturation Kit (Catalog #08571).
Subtype
Specialized Media
Cell Type
Neural Cells, PSC-Derived, Neural Stem and Progenitor Cells, Pluripotent Stem Cells
Species
Human
Application
Cell Culture, Differentiation, Organoid Culture
Brand
STEMdiff
Area of Interest
Disease Modeling, Neuroscience, Stem Cell Biology
Formulation
Serum-Free

Scientific Resources

Product Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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English
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English
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Safety Data Sheet 1
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Safety Data Sheet 2
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Safety Data Sheet 2
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08570
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English
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Safety Data Sheet 3
Catalog #
08570
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English
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Safety Data Sheet 4
Catalog #
08570
Lot #
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English
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Safety Data Sheet 5
Catalog #
08570
Lot #
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English
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Safety Data Sheet 6
Catalog #
08570
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All
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English
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Safety Data Sheet 7
Catalog #
08570
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English

Educational Materials (38)

Brochure

Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Figure 1.

(A) A representative phase-contrast image of a whole cerebral organoid at day 40 generated using the STEMdiff™ Cerebral Organoid Kit. 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). (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). (C-F) Inset of boxed region from (B). (C) PAX6+ apical progenitors (red, enclosed by dotted line) are localized to a ventricular zone-like region. β-tubulin III+ neurons (green) are adjacent to the ventricular zone. (D) CTIP2, a marker of the developing cortical plate, co-localizes with β-tubulin III+ neurons in a cortical plate-like region. Organization of the layers recapitulates early corticogenesis observed during human brain development. (E) Proliferating progenitor cells labeled by Ki-67 (green) localize along the ventricle, nuclei are counterstained with DAPI (blue). (F) An additional population of Ki-67+ cells is found in an outer subventricular zone-like region (arrowheads). Scale Bar = (A) 1 mm, (B) 500 µm and (C-F) 200 µm.

Figure 2.

Principal component analysis of hPSC and cerebral organoid transcriptomes. Cerebral organoids generated using the STEMdiff™ Cerebral Organoid Kit (filled blue circles) cluster together, and cluster with previously published (C Luo et al. Cell Rep, 2016) cerebral organoids (open blue circles). The first principal component accounts for the majority of variance seen (PC1; 80%) and distinguishes the cerebral organoid samples from the hPSCs (green circles). The second principal component accounts for only 9% of the variation, and highlights the modest expression differences between cultured organoids and primary embryonic fetal brain samples (19 post-conceptional weeks, brown circles).

Figure 3.

Heatmap of expression levels for genes associated with synaptic transmission function and neurogenesis in Day 40 organoids. These data show that gene expression of cerebral organoids generated from the STEMdiff™ Cerebral Organoid Kit are similar to published results (C Luo et al. Cell Rep, 2016).

Immunocytochemistry image of a cerebral organoid cultured in mTeSR™ Plus and directed to cerebral organoids using the STEMdiff™ Cerebral Organoid Kit.

Figure 4. Generation of Cerebral Organoids from hPSCs Maintained in mTeSR™ Plus

Human ES (H9) cells were cultured with mTeSR™ Plus and directed to cerebral organoids using the STEMdiff™ Cerebral Organoid Kit. Image shows apical progenitor marker SOX2 (purple) and neuronal marker TBR1 (green).

Publications (5)

One-Stop Microfluidic Assembly of Human Brain Organoids To Model Prenatal Cannabis Exposure. Z. Ao et al. Analytical chemistry 2020

Abstract

Prenatal cannabis exposure (PCE) influences human brain development, but it is challenging to model PCE using animals and current cell culture techniques. Here, we developed a one-stop microfluidic platform to assemble and culture human cerebral organoids from human embryonic stem cells (hESC) to investigate the effect of PCE on early human brain development. By incorporating perfusable culture chambers, air-liquid interface, and one-stop protocol, this microfluidic platform can simplify the fabrication procedure and produce a large number of organoids (169 organoids per 3.5 cm × 3.5 cm device area) without fusion, as compared with conventional fabrication methods. These one-stop microfluidic assembled cerebral organoids not only recapitulate early human brain structure, biology, and electrophysiology but also have minimal size variation and hypoxia. Under on-chip exposure to the psychoactive cannabinoid, $\Delta$-9-tetrahydrocannabinol (THC), cerebral organoids exhibited reduced neuronal maturation, downregulation of cannabinoid receptor type 1 (CB1) receptors, and impaired neurite outgrowth. Moreover, transient on-chip THC treatment also decreased spontaneous firing in these organoids. This one-stop microfluidic technique enables a simple, scalable, and repeatable organoid culture method that can be used not only for human brain organoids but also for many other human organoids including liver, kidney, retina, and tumor organoids. This technology could be widely used in modeling brain and other organ development, developmental disorders, developmental pharmacology and toxicology, and drug screening.
Human CNS barrier-forming organoids with cerebrospinal fluid production. L. Pellegrini et al. Science (New York, N.Y.) 2020

Abstract

Cerebrospinal fluid (CSF) is a vital liquid, providing nutrients and signaling molecules and clearing out toxic by-products from the brain. The CSF is produced by the choroid plexus (ChP), a protective epithelial barrier that also prevents free entry of toxic molecules or drugs from the blood. Here, we establish human ChP organoids with a selective barrier and CSF-like fluid secretion in self-contained compartments. We show that this in vitro barrier exhibits the same selectivity to small molecules as the ChP in vivo and that ChP-CSF organoids can predict central nervous system (CNS) permeability of new compounds. The transcriptomic and proteomic signatures of ChP-CSF organoids reveal a high degree of similarity to the ChP in vivo. Finally, the intersection of single-cell transcriptomics and proteomic analysis uncovers key human CSF components produced by previously unidentified specialized epithelial subtypes.
Cerebral organoids at the air–liquid interface generate diverse nerve tracts with functional output S. L. Giandomenico et al. Nature Neuroscience 2019 apr

Abstract

Neural organoids have the potential to improve our understanding of human brain development and neurological disorders. However, it remains to be seen whether these tissues can model circuit formation with functional neuronal output. Here we have adapted air–liquid interface culture to cerebral organoids, leading to improved neuronal survival and axon outgrowth. The resulting thick axon tracts display various morphologies, including long-range projection within and away from the organoid, growth-cone turning, and decussation. Single-cell RNA sequencing reveals various cortical neuronal identities, and retrograde tracing demonstrates tract morphologies that match proper molecular identities. These cultures exhibit active neuronal networks, and subcortical projecting tracts can innervate mouse spinal cord explants and evoke contractions of adjacent muscle in a manner dependent on intact organoid-derived innervating tracts. Overall, these results reveal a remarkable self-organization of corticofugal and callosal tracts with a functional output, providing new opportunities to examine relevant aspects of human CNS development and disease.

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