Induced Pluripotent Stem Cells (IPSCs) have the ability to turn into any cell in the body; however, IPSCs are very sensitive, and they can be difficult to grow in culture. While being cultured, IPSCs can easily suffer damage to their DNA which will lead to cell death. That limits their potential for use in treatments for diseases and conditions.
Now, researchers at the National Institutes of Health have found a way to protect stem cells from stress and maintain their normal stem cell structure. They developed a four-part small molecule cocktail which safeguards the cells and can be used to make stem cell use more efficient and predictable. The use of this small molecule cocktail not only prevents cell death, but it also improves the survivability and quality of stem cells which could help to significantly boost stem cell applications for research as well as clinical treatments.
The researchers believe their cocktail could help to advance the use of stem cells for treating diseases and conditions such as diabetes, Parkinson’s disease, and spinal cord injury and many more.
The small molecule cocktail – named CEPT – is described in a May 3 paper published in Nature Methods. The researchers explain their use of high-throughput screening to test thousands of drugs and compounds to identify unique combinations capable of improving stem cell survival by protecting against stress. The researchers tested more than 15,000 FDA-approved drugs and investigational small-molecule compounds from NCATS’ collections on IPSCs were generated from skin and blood cells.
The researchers tested 20 drugs that inhibited ROCK activity, a kinase enzyme involved in stem cell stress. Chroman 1 was found to improve survival to a much greater extent than the more widely used Y-27632. The researchers looked for drugs with potential synergies with Chroman 1 using NCATS’ matrix drug screening capabilities, which identifies possible interactions between different drugs. One investigational drug, Emricasan was found to further improve stem cell survival when combined with Chroman 1.
The researchers developed a new assay to replicate the stresses on stem cells during single cell cloning and examined the effects of over 7,500 compounds on 10 cells at a time. This new assay allowed them to identify a third compound, trans-ISRIB, that further improved cell survival, even when there were few cells in each plate. A mixture of polyamines was also found to improve survival when combined with Chroman 1, Emricasan and trans-ISRIB.
“Cells need to be cultured properly, and they have to be of good quality to go into patients,” said NCATS Acting Director Joni Rutter, Ph.D. “By finding new ways to protect stem cells from damage, these results could eventually have wide-ranging implications for many different diseases, including cancer, Alzheimer’s disease and more.”
Further experiments showed that CEPT improved the survivability of stem cells in cryopreservation, a process that is extremely stressful for the cells. Tests on IPSCs that had already been differentiated into mouse organ cells showed they were more viable with improved function when CEPT was used.
“Our approach could improve safety and ensure that the next-generation stem cell lines are cultured at high quality before moving into the clinic,” said Ilyas Singeç, M.D., Ph.D., director of the Stem Cell Translation Laboratory at NIH’s National Center for Advancing Translational Sciences (NCATS) and lead researcher of the study. “The cocktail will become a broadly used staple of the stem cell field and boost stem cell applications in both research and the clinic.”
You can read more about the study in the paper – A small-molecule cocktail that beats cellular stress – which was recently published in Nature Methods. DOI: 10.1038/s41592-021-01127-1