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A four-part small molecule cocktail can prevent cell stress

Researchers at the National Institutes of Health (NIH) have devised a four-part small molecule cocktail that can protect stem cells, including: Induced pluripotent stem cells (IPSC) Maintains normal stem cell structure and function from stress.

Researchers suggest that the cocktail may enhance potential therapeutic uses for stem cells, from treating diseases and conditions such as diabetes, Parkinson’s disease, and spinal cord injury to genome editing. ..

Human pluripotent stem cells are cells that theoretically grow forever and serve as an inexhaustible source of specialized cells such as the brain, kidneys, and heart. However, stem cells are sensitive and their potential use in medicine is hampered by the stress of growing in cell culture dishes, which can damage their DNA and lead to cell death.

In a series of experiments, Ilyas Singeç, MD, Ph.D., director of the Stem Cell Translation Institute at the National Center for Advancing Translational Sciences (NCATS) at NIH. Scientists led by systematically tested thousands of compounds using high-throughput screening. Drugs to identify unique combinations that significantly improve stem cell survival and reduce cell culture stress.

Singeç and his collaborators explained on May 3 how they developed a cocktail called CEPT and its potential uses. Nature method.

Small molecule cocktails protect cells and make stem cell use more predictable and efficient. It avoids cell death and improves the quality of surviving cells to prevent normally occurring cell stress and DNA damage. Cocktails have become a widely used staple food in the field of stem cells, supporting the application of stem cells in both research and clinics. “

Ilyas Singeç, MD, Ph.D., Director, Stem Cell Translation Laboratory, NIH’s National Center for Advancing Translational Sciences (NCATS)

iPS cells are derived from reprogrammed skin or blood cells. To improve their survival in culture, Singeç and his team first tested over 15,000 US Food and Drug Administration-approved drugs and small molecule compounds under investigation from the NCATS collection.

Among the 20 drugs and compounds that may inhibit the activity of ROCK, a kinase enzyme involved in stem cell stress, they are more likely to improve cell survival than the widely used compound Y27632. I found it to be powerful.

To further improve cell survival, Singeç and his colleagues used NCATS’s matrix drug screening feature to look for potential synergies between Chroman1 and other drugs and compounds. Matrix drug screening allows researchers to study the effects of drug combinations and determine the mechanisms by which these drugs may act.

Researchers have identified Emrikasan, an investigational drug that, when combined with Croman 1, can provide additional support for improving stem cell viability.

According to Singeç, an important effort in stem cell biology is an experimental process called single-cell cloning. Although it is easier to culture stem cells in large groups, single cell cloning (culturing one cell at a time in a small well on a cell culture plate) is very stressful and inefficient. is. This process has important uses for gene editing and establishment of cell lines, which are cell cultures developed from a single cell.

In the initial screening operation, the team tested the protective effect of the drug and compound on 500 stem cells at a time in platewells. To mimic the cellular stress seen during single-cell cloning, researchers could develop new assays (tests) to examine the effects of more than 7,500 compounds on just 10 cells at a time. It was.

This test identified trans-ISRIB, a third compound that enhances cell viability even when each plate is low in cells. Additional experiments have demonstrated that a mixture of compounds called polyamines (in combination with Chroman 1, Emricasan, and trans-ISRIB) is optimal for single-cell cloning.

“Cells need to be properly cultured and must be of good quality to enter the patient,” said Dr. Joni Latter, Deputy Director of NCATS. “By finding new ways to protect stem cells from damage, these results can ultimately have broad implications for many different diseases, including cancer, Alzheimer’s disease, and more.”

The team conducted a series of experiments to test the usefulness of the cocktail. Researchers have shown, for example, that CEPT improved biobanking of stem cells called cryopreservation. This involves freezing of the cells and is usually very stressful for the cells.

Cryopreservation is important for bringing stem cells to the clinic, but a significant number of cells are lost or damaged during the thawing process. Cocktails have dramatically improved the process.

In another test, researchers studied the use of cocktails in iPS cells that have already differentiated into heart cells, motor neurons, and other cell types.

They found that these more differentiated cells treated with CEPT were also more viable and showed improved function. Singeç also looked at the potential uses of cocktails in tissue engineering and the bioproduction of various cell types for regenerative medicine and drug development.

“For the last two decades, we haven’t been able to culture human stem cells under optimal conditions,” says Singeç. “Our approach has the potential to improve safety and ensure that next-generation stem cell lines are cultured in high quality before moving to the clinic.”

A four-part small molecule cocktail can prevent cell stress

Source link A four-part small molecule cocktail can prevent cell stress

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