WS6 enables scalable ex vivo expansion and gene editing of epithelial basal stem cells
Orr, J. C.; Haughey, E. K.; Farr, A. S.; Pearce, D. R.; McCarthy, N. A.; Reddy, S. K.; Rouhani, M. J.; Percival, C.; Rose, I.; Straatman-Iwanowska, A.; Dale, R.; Guthrie, M.; Benedetti, G.; Pape, O. R.; Ocampo-Godinez, J. M.; Maughan, E. F.; Butler, C. R.; Moulding, D. A.; Kreins, A. Y.; Giobbe, G. G.; De Coppi, P.; Grey, W.; Dost, A. F. M.; Hirst, R. A.; Baines, D. L.; Ishii, Y.; O'Callaghan, C.; Janes, S. M.; Hynds, R. E.
Show abstract
Modeling human epithelial diseases and developing cell-based therapies require robust methods to expand and manipulate epithelial stem and progenitor cells in vitro. Basal stem/progenitor cells from stratified epithelia can be expanded in 3T3-J2 fibroblast feeder cell co-culture systems, and the addition of the ROCK inhibitor Y-27632 enhances proliferation and culture longevity, a phenomenon described as conditional reprogramming. Here, we present a method incorporating the small molecule WS6 to further improve the proliferation and lifespan of cultured epithelial cells from multiple tissues, including airway, skin, and thymus. Cells maintained in this medium ( EpMED; FAD+Y+WS6) retain basal stem/progenitor cell identity and function, including the capacity to differentiate. We demonstrate their capacity to engraft in vivo in a tracheal transplantation model. In a second application, we generate clonal CRISPR-Cas9 genome edited nasal cultures, introducing targeted knockouts of DNAH5 or DNAI2 to create primary ciliary dyskinesia disease models. We anticipate that our method will have broad applications in epithelial cell biology, disease modeling, and regenerative medicine, while reducing reliance on immortalized or cancer cell lines and animal experimentation.
Matching journals
The top 7 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Structure-function relationships of mucociliary clearance in human and rat airways 96%
- Delayed induction of type I and III interferons mediates nasal epithelial cell permissiveness to SARS-CoV-2 95%
- Pre-ciliated tubal epithelial cells are prone to initiation of high-grade serous ovarian carcinoma 95%
Similar papers in this journal
Similar papers in this journal
Similar papers in this journal
- Integrin-activating Yersinia protein Invasin sustains long-term expansion of primary epithelial cells as 2D organoid sheets 96%
- Caspase-4/11 exacerbates disease severity in SARS-CoV-2 infection by promoting inflammation and thrombosis 94%
- MicroRNA-dependent inhibition of PFN2 orchestrates ERK activation and pluripotent state transitions by regulating endocytosis 94%
Similar papers in this journal
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.