STEMdiff Dorsal Forebrain Org Diff Kit

Generating dorsal forebrain organoids from human pluripotent stem cells (hPSCs), this matrix-free, serum-free differentiation system from STEMCELL Technologies completes guided patterning in 43 days through four stages: formation (Day 0-6), expansion (Day 6-25), differentiation (Day 25-43), and maintenance (Day 43+). When paired with AggreWell™800 plates for embryoid body aggregation, the kit supports scalable production of over 200 dorsal forebrain organoids per kit from lines such as H9 embryonic stem cells, each exhibiting layered cortical-like structures with glutamatergic neurons and progenitors expressing FOXG1, PAX6, TBR1, and CTIP2.

Components ship frozen as single use vials and bottles; store at -20°C, with select light-sensitive supplements held at -80°C. Thaw aliquots immediately before use, protect from light, and avoid repeated freeze-thaw cycles to preserve stability across the multi-week protocol. Researchers seed 3 x 10^6 cells per AggreWell™800 well at 10,000 cells per microwell, centrifuge plates at 1300 x g, and perform full-medium changes every two days on a 6-well plate under cell culture conditions at 37°C.

Validated for human-specific applications including disease modeling for microcephaly and autism spectrum disorder, neurodevelopmental studies, and high-content drug screening, the kit integrates upstream with mTeSR™1, mTeSR™ Plus, or TeSR™-E8™ maintenance media and downstream with the Neural Organoid Maintenance Kit. For research use only, the protocol yields reproducible in vitro models of the developing dorsal pallium for immunofluorescence characterization, RNA-seq phenotyping, and functional assays of human cortical development.