CultureSure™ CEPT Cocktail(1,000×)
CEPT is a cocktail of small molecules developed at the National Institutes of Health (NIH). It contains four components, and CEPT is an acronym for them: Chroman 1, Emricasan, Polyamines, and Trans-ISRIB.
In human ES cells and human iPS cells (human pluripotent stem cells [hPSCs]), CEPT prevents cellular stress and DNA damage during cell passage.1) Like Y-27632, a known potent ROCK inhibitor, it improves the viability of hPSCs during cell passage and cryopreservation.
Compared to existing methods, CEPT also improves cell viability in stem cell research, including embryoid body and organoid formation, single cell cloning, and genome editing using hPSCs.1-5)
Our product is a ready-to-use CEPT cocktail solution that has been tested for sterility and does not require preparation. This filter-sterilized solution can be added directly to the culture medium. It will aid in regenerative medicine research.
*This product is manufactured and sold under license from NIH.
Analytical Data
- Concentration (HPLC): Passed
- Appearance: Liquid
- Endotoxin: Less than 3 EU/mL
- Tested for sterility
- Tested for negative mycoplasma contamination
How to Use
Add 1/1,000 volume of this product to the culture medium and mix thoroughly before use.
Precautions for Use
- This product should be stored frozen (-20°C). To avoid repeated freezing and thawing, it is recommended to aliquot in small volumes and freeze.
- Some precipitates may be visible when dissolving. This will not affect the quality of the product, and it can be used as it is.
(It is possible to dissolve the precipitates by incubating at 37°C for 1~2 hours, but repeated freezing and heating should be avoided.)
Samples and Applications
Samples: human ES cells, human iPS cells
Applications:
Data
Colony Formation Test
The human iPS cell 201B7 strain was seeded in StemSure® hPSC Medium Δ containing bFGF supplemented with CEPT or Y-27632 at 37°C under 5% CO2 conditions (6-well plates, 1 well= 9.5 cm2).
After at least 16 hours from the start of culture, the medium was replaced with the medium without CEPT or Y-27632, and the culture was continued with media changes every 2 to 3 days.
After checking the cells on day 8, they were fixed with 4% paraformaldehyde, stained with ALP, and colonies were counted.
Single-cell Cloning Test
The human iPS cell 201B7 strain was seeded in StemSure® hPSC Medium Δ containing bFGF supplemented with CEPT or Y-27632 at 37°C under 5% CO2 conditions (96-well plates).
After at least 16 hours from the start of culture, the medium was replaced with the medium without CEPT or Y-27632, and the culture was continued with media changes every 2 to 3 days.
After checking the cells on day 13, they were fixed with 4% paraformaldehyde, stained with ALP, and the number of wells with and without colonies were counted. The colony formation rate was calculated from the number of wells with colonies relative to the total number of wells.
[Result]
When the number of cells seeded was small, more colonies were formed in the CEPT-supplemented cultures than in the Y-27632-supplemented cultures.
Undifferentiated State Maintenance
- Cells: human iPS cell 201B7 strain
- Medium: StemSure® hPSC Medium Δ + 35 ng/mL bFGF
- Red: rBC2LCN-635 (human iPSC membrane stain)
- Blue: DAPI (nuclear stain)
- Note: BC2LCN is a recombinant lectin with high affinity for cell surface glycans of human ES cells and human iPS cells.
-
▽With CEPT 
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▽With Y-27632 
[Result]
No difference in cell morphology was observed between the addition of CEPT and Y-27632. The undifferentiated state was also maintained.
References
- Chen, Y. et al. : Nat. Methods, 18(5), 528 (2021).
A versatile polypharmacology platform promotes cytoprotection and viability of human pluripotent and differentiated cells. - Tristan, C.A. et al. : Stem Cell Reports, 16(12), 3076 (2021).
Robotic high-throughput biomanufacturing and functional differentiation of human pluripotent stem cells. - Tristan, C.A. et al. : Nat. Protoc., 18(1), 58 (2022).
Efficient and safe single-cell cloning of human pluripotent stem cells using the CEPT cocktail. - Deng, T. et al. : Stem Cell Reports, 18(4), 1030 (2023).
Scalable generation of sensory neurons from human pluripotent stem cells. - Takeshi, W. et al. : Regen. Med., 18(3), 219 (2023).
International evaluation study of a highly efficient culture assay for detection of residual human pluripotent stem cells in cell therapies.
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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