Technical Notes
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Human pluripotent stem cell (hPSCs) are notoriously sensitive cells that undergo a unique form of dissociation-induced apoptosis. When passaging hPSCs as single cells, cellular stress and impairment of cell structure are seen immediately upon dissociation – and are most noticeable for several hours after.
In the groundbreaking 2021 paper from Chen et al., researchers identified three small molecules consisting of Chroman 1, Emricasan, and Trans-ISRIB (CET) combined with a polyamine solution to create the CEPT supplementation strategy, or a CEPT cocktail solution, which was shown to provide comprehensive cytoprotection to effectively promote cell attachment and survival of dissociated cells.
Figure 1: Superior cell survival with CEPT. hPSCs seeded as single cells were incubated with Y27632 ROCK inhibitor and various small molecule combinations of the CEPT cocktail, highlighting combinatorial improvements to cell survival. Alkaline Phosphatase (AP) staining shows visually the increased number of colonies and growth in clonal cultures treated with CEPT over Y27632 or the commercially available CloneR supplement. Published data from Yu Chen et al. (2021) Nature Methods.
Supplementation with CEPT during dissociation steps rapidly reduces cell blebbing (an indicator of a cell undergoing late-stage apoptosis) and maintains cell function and integrity. Additionally, CEPT promotes more efficient cell adhesion to the culture substrate. From routine cell passaging to embryoid body (EB) differentiation, to organoid formation, to single-cell cloning and gene editing, CEPT can be used in a variety of applications and various cell types.
In 2021, Captivate Bio set out to bring this new hPSC survival supplement to market in a product format, aligning with our long history of commercializing small molecules and other key tools for stem cell research. With the launch of our CET Cocktail, a kit composed of the key small molecules Chroman 1, Emricasan, and Trans-ISRIB, we are the first to offer this small molecule pack to researchers seeking to improve the viability and functionality of their stem cell cultures.
The CET Cocktail provides:
Captivate Bio is committed to supporting our customers with new technology to speed discovery. We are actively recruiting labs in validating CET through our Collaboration Program, contact us today to learn more.
Learn more about how we support researchers with cell culture products, contract manufacturing and storage services, custom reagent sourcing, as well as marketing, sales, and distribution programs for our industry partners.
The CEPT cocktail is a ground-breaking supplementation strategy for improving hPSC viability and survival under stressful conditions. Read more to learn how CEPT can keep your cells happy with a chemical hug.
This Research Spotlight highlights a recent publication showing a reproducible and scalable workflow for establishing clonal cell lines and a standardized gene editing procedure using the CEPT Cocktail.
This Research Spotlight highlights the CEPT cocktail, a chemically defined molecule combination that has been shown to provide superior cell survival under stressful conditions, such as single-cell passaging, cloning, and gene editing protocols.
