STEMdiff™ Cerebral Organoid Kit
Culture medium kit for establishment and maturation of human cerebral organoids
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Y-27632 (Dihydrochloride)RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
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Ultra-Low Adherent Plate for Suspension Cultu...Sterile, flat-bottom plate, with ultra-low adherent surface for suspension cultures, with lid; 6-well format
What Our Scientist Says
Human brain development is complex, so to observe it in a dish is a huge scientific breakthrough. We've tried to simplify that process and make it more accessible to you, regardless of how much stem cell experience you have.
Overview
These defined, serum-free cell culture media and the 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.
Data Figures
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).
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).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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.
Resources and Publications
Educational Materials (38)
Publications (5)
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Legal Statement:
This product was developed under a license to intellectual property owned by the Institute of Molecular Biotechnology (IMBA) of the Austrian Academy of Sciences. This product is sold for research use only (whether the buyer is an academic or for-profit entity) under a non-transferable, limited-use license. Purchase of this product does not include the right to sell, use or otherwise transfer this product for commercial purposes (i.e., any activity undertaken for consideration, such as use of this product for manufacturing, or resale of this product or any materials made using this product, or use of this product or any materials made using this product to provide services or, in collaboration with, a for-profit entity, for purposes other than research applications (i.e., drug development activities). Purchasers wishing to use the product for commercial purposes should contact IMBA at technologvtransfer@imba.oeaw.ac.at.
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
