The human cerebral organoids are 3D neural tissues derived from human pluripotent stem cells. They reproduce key features of the developing brain. The STEMdiff™ Cerebral Organoid Kit from StemCell Technologies offers a defined, serum-free system with a clear four-stage process for consistent organoid generation. This protocol follows the method described by Lancaster et al. (Nature 2013; Science 2014). It guides cells through embryoid body formation, neuroepithelial induction, Matrigel embedding, and maturation. Scientists in any laboratory can achieve reproducible brain-like organoids with minimal variability.
Kit Composition and Protocol
The kit contains all media required for each culture stage. It includes two basal media and five supplements. These components combine into four complete formulations—one for each stage of the protocol.
Researchers start by dissociating pluripotent stem cells to form embryoid bodies during days 1–5. Induction medium is applied on days 6–7. EBs are embedded in Matrigel and expanded from days 7–10. Long-term maturation occurs from day 10 onward with gentle agitation. The serum-free design eliminates variability and ensures reliable results. This straightforward approach works well in South African laboratory environments equipped for standard cell culture.
Cortical Organoid Architecture
Organoids generated with this kit develop layered neural regions resembling the human cortex. By day 40, they typically show:
- Ventricular zones with SOX2^+, PAX6^+, and Ki‑67^+ progenitors
- Subventricular zones with Ki‑67^+/p-Vimentin^+ cells
- Intermediate zones rich in TBR2^+ progenitors
- Cortical plates containing CTIP2^+, MAP2^+, and TBR1^+ neurons
These structures form in orientations similar to those found in vivo. Researchers can observe neuroepithelial folds under phase-contrast microscopy and confirm marker expression via immunostaining. The forebrain-like identity (including FOXG1 expression) makes these organoids suitable for disease modeling and neurodevelopment studies.
Extended Culture and Maturation
Cultures exceeding 40 days benefit from additional supplements. The STEMdiff cerebral organoid maturation kit (sold separately) supports further neuronal development and synapse formation. This enables long-term studies requiring more mature neurons, such as electrophysiology or advanced imaging.
Existing laboratory equipment—such as orbital shakers or bioreactors—can maintain nutrient exchange without additional investment.
Key Features:
- Defined, serum-free media (2 basal + 5 supplements)
- Four-stage protocol: EB formation → induction → Matrigel expansion → maturation
- Produces layered cortical-like organoids with distinct brain regions
- Reliable results for neuroscience and stem cell research
Applications and Apex Scientific Support
The kit supports applications in disease modeling, drug discovery, neurodevelopment, and neurotoxicity testing. It integrates smoothly with assays including immunofluorescence, phenotyping, and functional analysis.
Apex Scientific delivers the STEMdiff cerebral organoid kit across South Africa and advises on complementary laboratory equipment. Their technical team helps researchers optimise workflows for organoid culture and related neuroscience applications. For more insights, see the Apex Scientific Insights page.
