STEMdiff Branching Lung Organoid Kit

From STEMCELL Technologies, this serum-free medium system supports the efficient and reproducible generation of branching lung organoids from human embryonic stem (ES) and induced pluripotent stem (iPS) cells. The kit guides cells through four sequential stages of differentiation — definitive endoderm, anterior foregut endoderm, lung bud organoid, and branching lung organoids — and has been optimized for differentiation of cells maintained in mTeSR™1.

Resulting organoids develop proximal and distal-like branching airway epithelial structures expressing EPCAM, NKX2.1, SOX2, SOX9, MUC1, and P63. The workflow progresses with stage 1 complete by day 3 (definitive endoderm), anterior foregut endoderm buds released during stage 2 (days 3–6), ventralized lung bud organoids forming during stage 3 (days 6–14), and lung bud organoids embedded in a Matrigel® sandwich during stage 4 to mature into branching lung organoids. When maintained beyond 28 days in a Matrigel® sandwich, expression levels of markers for more mature lung cells, such as SFTPC, SFTPB, and ABCA3, increase. Organoids generated in the kit also express proteins associated with SARS-CoV-2 entry, ACE2 and TMPRSS2.

The kit supports long-term culture, with NKX2.1 and CPM expression maintained in cultures to day 67 and branching tips continuing to grow up to day 101. For extended organoid culture beyond 28 days, additional components for branching lung organoid maturation are available in a separate STEMdiff™ Branching Lung Organoid Maturation Kit. All components are sold as part of the kit and are not available for individual sale, and the product is intended for research use only.