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Highly potent and selective ROCK inhibitor with greater effectiveness at lower doses than Y27632. Enhances cell survival of dissociated hPSCs.
Human pluripotent stem cells (hPSCs) are extremely sensitive to environmental changes, and require safe, controlled, and efficient strategies for the development, expansion, and banking of cell-based therapeutic products. Chroman 1 is a highly potent and selective ROCK inhibitor (both ROCK 1 and ROCK 2) that promotes survival of dissociated hPSCs in culture when used at a final concentration of 50 nM. Significant improvements to cell viability, and expansion are seen when Chroman 1 is applied in combination with Emricasan, Polyamines, and Trans-ISRIB, a combination known as CEPT.
Chroman 1 is more effective at lower doses than Y27632, with ROCK1 IC50 = 52 pM and ROCK2 IC50 = 1 pM.
Chroman 1 is also more specific than Y27632, not showing any significant kinase inhibition outside ROCK 1/2 at 50 nM, unlike off-target effects observed with Y27632 applied at 10 µM1. Chroman 1 exhibits > 2000-fold selectivity for ROCK 2 over a range of related kinases including MRCK (IC50 = 150 nM), PKA (IC50 >20000 nM), and AKT1 (IC50 >20000 nM). Chroman 1 is a component of Captivate Bio's CET Cocktail.
Alternative names: ROCK-II Inhibitor, ROCK2 Inhibitor Applications: Pluripotent Stem Cell Culture; Single-Cell Cloning; Gene Editing; Cryopreservation & Cell Banking; Tissue Engineering; Biomanufacturing; Regenerative Medicine Cell types: Neurons, Pluripotent Stem Cells, iPSCs, ESCs, organoids
The data provided herein may relate to products or workflows that have not been fully validated by Captivate Bio and is subject to change without notice.
References and sample data are from publications that support the biological activity of the product. This data reference is from Chen et al. 2021 Nat Methods.
Figure 1. The potency Chroman 1 was assessed against ROCK1 and ROCK2 showing high potency of Chroman 1 and relative IC50 results for each ROCK target.
Figure 2. Improved cell survival was seen with dissociated cells passaged with Chroman 1, as seen by phase-contrast and fluorescent images depicting stained live and dead cells taken 12 hours after plating.
Figure 3. hPSCs cultured for 40 passages with Chroman 1 maintained a normal karyotype, expressed expected pluripotency markers, and efficiently differentiated into ectoderm, mesoderm, and endoderm through directed differentiation protocols and embryoid body formation.
Keywords: Chroman 1, Chroman 1 supplier, Selective, p160, ROCK, inhibitors, inhibits, Rho-Kinase, Rho-Associated, Coiled-Coil, Kinases, ROK, stem, cells, organoids, iPSC, ROCKI, ROCK1, ROCKII, ROCK2, proliferation, reprogramming, cryopreservation, differentiation, cell freezing media, cryogenesis, self-renewal, organoids, neural, Y-27632
Prepare a concentrated stock solution of Chroman 1 by reconstituting the lyophilized powder with DMSO to the appropriate stock concentration. Generally a 1,000X or 10,000X concentrated stock solution is useful, depending on the desired final concentration and application. Refer to the Chroman 1 Technical Data Sheet and/or the CET Cocktail product flier for more details on the reconstitution and use of the small molecule for stem cell applications.
Chroman 1 is more potent and selective than other commonly used ROCK inhibitors, including Y27632 and Thiazovin. Chroman 1 has a greater effectiveness at lower doses than Y27632.
When used to enhance hPSC survival and clonal expansion during passaging, Chroman 1 is used at a final concentration of 50 nM in cell culture media. Chroman 1 is generally added to culture media at passaging and is removed during the first feeding the following day (24 hour application).
Chroman 1 is soluble in DMSO to 50 mg/mL (approx. 115 mM), with an unknown saturation point.
Chroman 1 is a potent inhibitor of ROCK (rho-associated coiled-coil kinase). Chroman 1 affects the Wnt, TGF-beta/Smad pathways and acts as a cytoprotectant for plurpotent stem cells under stress. Chroman 1 is also an effective inhibitor of MRCK.
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