At Cell2in, we are continuing to develop our cutting-edge technology, FreSHtracer, through cell therapy.
Cells separated according to their GSH levels perform differently. GSH-High cells were able to recover their GSH faster after oxidant treatment.
GSH-High Cells were also more functional in other areas of cellular therapy listed below.
The Benefit of Using FreSHtracerTM with Stem Cells
Some of the basic properties of stem cells are that they can self-replicate and migrate to a damaged area in the body to help repair it. When we tested the difference between stem cells with varying GSH levels, we found GSH-High stem cells outperformed all other stem cells.
- After using FreSHtracerTM and FACS sorting the cells, the GSH-High stem cells (hBM-MSC) grew and formed about 50% more colonies than GSH-Low stem cells and about 33% more than the unsorted stem cells.
- Again, after using FreSHtracerTM to measure GSH levels and sort stem cells with a FACS machine, we tested their migration response to a chemical attractant. After 24 hours, the GSH-High stem cells with were found in higher numbers in the migration tests. GSH-High stem cells are faster than all other stem cells.
GSH-High stems cells have increased stemness, they can not only self-replicate faster but can also migrate faster.
Asthma Treatment with FreSHtracer Stem Cells
Currently, there is no cure for asthma and can only be managed by avoiding allergens and irritants, and use of inhaled corticosteroids. Currently, about 250 million people in the world are suffering from asthma.
Recently, stem cell therapy has shown to be a promising therapy for alleviating the symptoms of asthma. So, at Cell2in, we decided to test our GSH-High stem cells to see if we could discover better results.
In a mouse model, we simulated a viral infection, using Poly I:C, to provoke the immune system in the lungs causing inflammation – similar to the inflamed lungs in asthma. We repeatedly provoked the immune system throughout the 24-day test. On day 14 of the test, we injected our mice intravenously with stem cells. Until the 24th day, the immune system was provoked several times more.
Below in the pictures: Bronchioles (where air passages through the lungs) are show as the circles. Inflammation is show as purple around the bronchioles.
- The lungs were not provoked and the bronchioles show no inflammation (a clean pink color).
- Lungs were provoked, and no stem cells were injected in this test. Note the large amount of inflammation.
- The unsorted stem cells were able to reduce some of the inflammation.
- With GSHLow stem cells, some reduction of inflammation, similar to the unsorted stem cells, can be seen.
- Our GSHHigh stem cells showed a remarkable reduction in inflammation similar to our unprovoked control test.
GSHHigh stem cells that were sorted with FreSHtracerTM, performed better at reducing inflammation in the lungs.
To see the other benefits of GSH-High stem cells in asthma treatment, see our academic paper:
The Power of GSHHigh Neurons
GSH depletion and GSH-related enzyme deficiency is thought to play key roles in several brain diseases including Autism, Alzheimer’s disease, bipolar disorder, Parkinson’s disease, and schizophrenia.
We tested mouse embryonic stem cells (mESC) that had been FreSHtracerTM sorted into categories of High, Low, and Unsorted stem cells(Naive). We then observed dendrite formation after the stem cells had differentiated into neurons. Dendrites, important for communication in the brain, are the short branches which receive synapses from other cells and transmit impulses to the rest of the neuron.
In the Naive stem cells, only a small amount of dendritic growth was exhibited.
In the three tests performed with GSHLow stem cells, dendrite growth remained low.
GSHHigh stem cells which different into neurons have higher numbers of dendrites. GSHHigh stem cells might prove effective at one day treating brain disorders.
Antioxidant Screening / Food and Drug Screening / Cosmetic Screening with FreSHtracerTM
If GSH-High cells are more functional and viable, then through compounds, it is possible to create more GSH-High cells by boosting the amount cellular GSH or increasing the rate of GSH Recovery Capacity (GRC).
After incubating the cells for 24 hours with an antioxidant, a pharmaceutical drug, food, or a cosmetic compound, we can then introduce an oxidizing agent – which will consume some or all of the active GSH in the cell – to measure how fast the cell can recover its GSH back to normal levels.
- FreSHtracerTM bound to GSH (F510) will show a bright green fluorescence. When an oxidizing agent has been added, FreSHtracerTM will no long be able to bind to GSH(Oxidized GSSG), and the cells lose their green fluorescence. As the the cells regenerate their GSH, FreSHtracerTM is able to bind again with GSH and the cells will get brighter in their green fluorescence.
- We measure the rate of GSH recovery following the addition of an oxidizing agent. Any compound that increases the rate of recovery in GSH has a beneficial effect for cells. In this test, Compound B was effected less by the oxidant and was able to recover GSH to a high level, 14% better than our control. We then take these GSH improving compounds, like Compound B, and add them to cell culture media or cosmetics to improve the performance of those cells.