Summary

U-Factor Co., Ltd. (hereinafter referred to as “U-Factor”) has collaborated with Senior Principal Researcher Yuji Teramura from the Department of Cell Molecular Engineering at the National Institute of Advanced Industrial Science and Technology (AIST) to discover that purified components from immortalized human deciduous tooth-derived stem cells (SHED) enhance cellular activity, improve cell migration ability, and retain antioxidant activity. These findings indicate that the technology holds the effects of stem cell transplantation, suggesting that it could be used as a safer and more efficient alternative to stem cell transplantation.

The details of this technology were published in “Scientific Reports” on July 3, 2024.

Social Background of the Development

In recent years, treatments using stem cells have gained attention in the field of regenerative medicine. However, stem cell transplantation carries risks of immune rejection and tumor formation, making it challenging to balance safety and efficacy. As a result, there is a growing demand for new treatments using culture supernatants derived from stem cells. These supernatants contain multiple cytokines and are expected to promote cell regeneration and repair. However, there are many unexplored aspects regarding their safety, quality, and efficacy, especially when using primary cultured cells, which have limitations in cell proliferation, production of supernatants, and the presence of impurities. Overcoming these challenges to develop safe and effective culture supernatants has become an urgent task.

Research Background

Professor Emeritus Minoru Ueda of Nagoya University Graduate School of Medicine, a director at U-Factor, has been conducting research on culture supernatants containing cytokines derived from stem cells for many years (K Matsubara, M Ueda, et al. J. Neurosci 2015). This research confirmed the efficacy of culture supernatants in treating spinal cord injuries and acute organ injuries. However, using primary cultured cells posed limitations in cell proliferation and the production of culture supernatants. To address these issues, we aimed to achieve a more stable supply and higher quality culture supernatants using immortalized deciduous tooth-derived pulp stem cells.

Research Details

This study evaluated the effects of the raw culture supernatant from immortalized deciduous tooth-derived pulp stem cells (hereinafter referred to as “raw supernatant”) and its extracted components (hereinafter referred to as “extracted components”) on various cells. We compared the raw supernatant and extracted components in terms of intracellular dehydrogenase activity, migration activity, and antioxidative stress activity. The extracted components showed equal or higher activity than the raw supernatant from primary cultured cells.

1. Dehydrogenase Activity Experiment

In this experiment, we assessed the effects of the culture supernatant by adding it to mouse fibroblasts (NIH3T3 cells). The results showed that the extracted components (red line) had higher cell activity compared to the raw supernatant (black line) and other extracted components (blue and yellow lines). The graph on the right shows similar results for the raw supernatant, confirming the findings from primary cultured cells.

2. Scratch Assay Experiment

This experiment evaluated cell migration ability (motility) by creating a scratch on a cell layer and observing how the cells filled the wound after adding the raw supernatant or extracted components. The cell recovery area (%) was used to evaluate how effectively the supernatants promoted cell migration and repair. The extracted components demonstrated equivalent cell migration ability compared to the raw supernatant (CM).

3. Oxidative Stress Experiment

This experiment assessed cellular resistance to oxidative stress by evaluating cell damage caused by hydrogen peroxide treatment. The results showed that the raw supernatant and extracted components 1 could suppress cell damage caused by hydrogen peroxide, while other extracted components did not demonstrate this effect. This confirms that the raw supernatant and extracted components effectively suppressed cell damage due to oxidative stress.

Furthermore, the extracted components successfully removed impurities such as ammonia and lactic acid while retaining a high concentration of cytokines (VEGF, HGF, bFGF) believed to be active ingredients. This purification resulted in a high-quality supernatant with active ingredients that function effectively, significantly improving treatment safety and efficacy.

Future Plans

Moving forward, we will continue to develop concrete treatments using the extracted components. We aim to innovate further in the field of regenerative medicine and offer new treatment options that improve patients’ quality of life. Through ongoing research and clinical trials, we will verify safety and efficacy and take steps towards practical application. Ultimately, we aim to introduce our technology into clinical practice as soon as possible, benefiting many patients.

Paper Information

Journal: Scientific Reports
Publication Date: July 3, 2024
Title: Activation of cellular antioxidative stress and migration activities by purified components from immortalized stem cells from human exfoliated deciduous teeth
Authors: Yujing Shu, Masato Otake, Yasuhiro Seta, Keigo Hori, Akiko Kuramochi, Yoshio Ohba, Yuji Teramura
DOI: 10.1038/s41598-024-66213-8

Glossary

SHED (Stem cells from Human Exfoliated Deciduous teeth)
Stem cells extracted from deciduous teeth. These cells have high proliferation and differentiation potential and are gaining attention in regenerative medicine and tissue repair research. SHED can differentiate into various cell types, including nerves, bones, and muscles, offering potential applications in diverse treatments in the future.

Immortalization
A technique that enables cells to proliferate indefinitely without undergoing natural aging or death. Normally, cells lose function and eventually die after a certain number of divisions. However, with immortalization technology, cells can avoid this aging process, allowing continuous division and proliferation. This technology is highly valuable in research and medicine, particularly in regenerative medicine and cancer research.