Introduction
To advance biomedical research, the culture and maintenance of stem cells is both a source of opportunities and a formidable challenge to carry out. Stem cell-based approaches hold the promise of advancing basic research, regenerative therapies, and personalized medicine. At Defined Bioscience, we produce fully defined products for researchers that enhance the stability, reproducibility, viability, and functionality of cultured stem cells. One of our latest products, Ready-CEPT cocktail for viability enhancement, advances this mission.
The origin of Ready-CEPT
The journey of Ready-CEPT began at the National Institutes of Health (NIH), where the need for a robust method to improve stem cell survival and health was identified. The traditional processes of passaging, cryopreservation, and thawing often expose stem cells to stress, leading to decreased viability, genetic shift, and compromised functionality. Recognizing these challenges, NIH researchers developed an additive cocktail that would not only support stem cells through these critical steps, but also enhance their overall health and genomic stability.
After substantial research and development, the CEPT cocktail emerged as the best in class formulation to support cell viability and cell line stability under several challenges. The name 'CEPT' encapsulates the essence of this innovation—Chroman 1 (ROCKi), Emricasan (casp3), Polyamines (anoikis), trans-ISRIB (integrated stress response). Scientific studies, including those published in Nature Methods1,2 and Nature Protocols3, have validated the effectiveness of CEPT, demonstrating its superiority in facilitating single-cell cloning, sorting, and improving cell survival rates.
What is Ready-CEPT?
Ready-CEPT is a fully-defined cocktail composed of four key components:
- Chroman-1, a ROCK inhibitor
- Emricasan, a pan-caspase inhibitor
- Polyamines including putrescine, spermine, and spermidine
- Trans-ISRIB, an integrated stress response inhibitor
Each of these components plays a vital role:
Chroman-1 mitigates the effects of enzymatic dissociation and mechanical handling, thereby reducing cell stress and death
Emricasan protects cells from apoptosis triggered during the freezing and thawing processes
Polyamines are essential for cellular functions such as DNA stabilization and protein synthesis, which are crucial during cell division and growth as well as prevent anoikis for adherence-dependent cells in suspension
trans-ISRIB supports protein homeostasis, enhancing the cells' ability to recover from stress
This multifunctional blend not only supports the cells during routine laboratory processes but also enhances their overall growth and stability, making it particularly beneficial for maintaining genetically stable stem cell cultures.
What does Ready-CEPT mean for your stem cell culture?
At Defined Bioscience, we understand that the needs of cell biologists are as diverse as the cells they study. Ready-CEPT cocktail is designed to support a variety of research settings, offering flexibility and compatibility with different cell culture media, including our specially formulated HiDef-B8 medium supplement and HiDef-B8 Complete that includes DMEM/F12.
Ease of Use: Ready-CEPT is provided as a 1000X concentrate in a user-friendly liquid form, making it easy to add to any culture medium without the need for extensive preparation or measuring out several components separately. This ease of use ensures that researchers can spend more time on their scientific inquiries and less on routine cell reagent preparation.
Quality and Reliability: Each batch of Ready-CEPT undergoes rigorous performance testing with human pluripotent stem cells (PSCs) to ensure consistent and reliable results. Our commitment to quality means that researchers can trust in the performance of Ready-CEPT to maintain the health and viability of their valuable cell cultures.
Enhancing stem cell outcomes
The application of Ready-CEPT in stem cell research has led to notable enhancements in cell viability across various procedures while maintaining functionality:
Cryopreservation and Thawing: Ready-CEPT significantly improves cell recovery rates post-thaw, maintaining cell integrity and functionality.
Passaging: It facilitates smoother transitions during passaging and subculturing, reducing cell death and enhancing colony formation.
Experimental Flexibility: The robust nature of Ready-CEPT treated cells enables more consistent results in downstream applications such as gene editing, organoid formation, and disease modeling.
Tips for optimal use
To maximize the benefits of Ready-CEPT, we recommend the following practices:
Proper Storage and Handling: Store Ready-CEPT at -20°C and protect it from light to preserve its efficacy. Handle the product with care, maintaining sterile conditions to avoid contamination.
Application Specifics: Before use, allow the cocktail to warm to room temperature. When adding to your medium, ensure that the final concentration is correctly adjusted for the specific needs of your cell type.
Conclusion
At Defined Bioscience, our mission is to advance the field of stem cell research by providing defined solutions like Ready-CEPT. By improving the survival, health, and functionality of stem cells, we enable researchers to push the boundaries of what is possible in biomedical research—at lower cost and with less labor. Whether for basic research, disease modeling, or therapeutic development, Ready-CEPT cocktail is a helpful tool for cultivating success in stem cell culture.
We invite researchers and biologists to experience the transformative effects of Ready-CEPT in their stem cell cultures. Together, we can unlock the full potential of stem cell research and pave the way for groundbreaking advances in medicine and biology.
References
- Pal, R. A small-molecule cocktail that beats cellular stress. Nat Methods 18, 457–458 (2021). https://doi.org/10.1038/s41592-021-01127-1
- Chen, Y., Tristan, C.A., Chen, L. et al. A versatile polypharmacology platform promotes cytoprotection and viability of human pluripotent and differentiated cells. Nat Methods 18, 528–541 (2021). https://doi.org/10.1038/s41592-021-01126-2
- Tristan, C.A., Hong, H., Jethmalani, Y. et al. Efficient and safe single-cell cloning of human pluripotent stem cells using the CEPT cocktail. Nat Protoc 18, 58–80 (2023). https://doi.org/10.1038/s41596-022-00753-z
- Ryu, S. et al. Stress-free cell aggregation by using the CEPT cocktail enhances embryoid body and organoid fitness. Biofabrication 16(1) (2023). https://doi.org/10.1088/1758-5090/ad0d13
- Ready-CEPT Product Guide