May 8, 2020

Research field: life science and medicine; Keywords: antiperspirant, heat stroke, hyperhidrosis, immortalized cell,long-term culture, myoepithelial cell, regenerative medicine, sweat gland

Applicable to the development of next-generation antiperspirants and to research on sweating dysfunction
and sweat gland regeneration

Successful generation of immortalized human eccrine sweat
gland myoepithelial cells—Concluded a sales license agreement with Applied Biological Materials Inc. in Canada—

Gist of the researchWe successfully generated immortalized human eccrine sweat gland myoepithelial (iEM) cells that retain thecharacters of myoepithelial cells.
The generated cell line will be sold globally through Applied Biological Materials Inc.
This achievement will help advance research on sweat glands, which will lead to new approaches in thedevelopment of antiperspirants, development of methods to prevent and treat sweating dysfunctions suchas hyperhidrosis and heat stroke, and regeneration of sweat gland tissue.

Overview
Our research group, led by Guest Professor Fumihiro Okada of the Laboratory of Advanced Cosmetic Science ※ 1,Graduate School and School of Pharmaceutical Science, Osaka University and Professor Kiyotoshi Sekiguchi of theDepartment of Contribution Research, Institute for Protein Research, Osaka University, has succeeded in generating
immortalized※2 human eccrine sweat gland myoepithelialcells※3.
Myoepithelial cells are one of the several types of cellthat make up human eccrine sweat glands. They areinvolved in the contraction of sweat glands and function asstem cells tomaintain the function of sweat glands (Fig. 1). Identificationof the components that regulate the function of humaneccrine sweat gland myoepithelial cells will lead to new
approaches in the development of antiperspirants and tomethods of prevention and treatment of hyperhidrosis andheat stroke. To achieve these goals, we need many humaneccrine sweat gland myoepithelial cells. However, up until
Figure 1.
now, no human eccrine sweat gland cell line have retained
Structure of the human eccrine sweat glandthe characters of human eccrine sweat gland myoepithelialcells.
Our research group had earlier established a method for culturing human eccrine sweat gland myoepithelial cells
such that they retained their functional characters for a short period of time (refer to the News Release of October 27,2016, https://www.mandom.co.jp/release/pdf/2016102701.pdf [in Japanese]). This time, by using this methodtogether with the introduction of an immortalizing gene, we have achieved a breakthrough in generating immortalized
human eccrine sweat gland myoepithelial (iEM) cells that can be cultured for a long time.
This research achievement will accelerate research in this field, which will lead to the development of nextgeneration antiperspirants, the early establishment of methods for prevention and treatment of hyperhidrosis andheat stroke, and the establishment of techniques for regenerating sweat glands.
We have filed a patent application on the generation of iEM cells and we presented our research at the 49th Annual
European Society for Dermatological Research Meeting in Bordeaux, France, 18–21 September 2019. The usefulnessof iEM cells was accepted, and Mandom corporation (headquartered in Osaka city, Osaka, Japan. President: MotonobuNishimura) concluded a license agreement with Applied Biological Materials Inc. (headquartered in Richmond, BritishColumbia, Canada)※4 for consignment sales of iEM cells. Additionally, Mandom corporation concluded a memorandumof understanding royalty with Osaka university (headquartered in Suita city, Osaka, Japan. President: Shojiro Nishio).
This material transfer agreement (MTA) ※5 has made it possible to transfer iEM cells to many countries worldwide.
Product HP: https://www.abmgood.com/immortalized-sweat-gland-myoepithelial-cells.html#T0816

Background of the research
Solving sweat-related concerns that occur in daily life, for example, hyperhidrosis and sweat smell, can improve
people’s quality of life. We thus focused on the regulation of sweating. To control sweating, we need to understandthe characteristics of the cells that make up sweat glands and regulate the function of sweat glands. Myoepithelial
cells, one of the various types of cell that make up human eccrine sweat glands, have two functions: they contract
sweat glands to push sweat out of the body surface and they function as stem cells to maintain the homeostasis of thesweat glands (Fig. 1). Therefore, the activity of myoepithelial cells plays an important role in human sweating. Culturedcells are a useful means for elucidating the functions of human eccrine sweat gland myoepithelial cells; however, nomethod had been available for their long-term culture while retaining the characters of human eccrine sweat gland
myoepithelial cells.

Research content
To date, cultured human eccrine sweat gland cells have notretained the characters of myoepithelial cells. Our researchgroup earlier established a method enabling human eccrine
sweat gland cells to maintain the characters of myoepithelial
cells for a short time by culturing human eccrine sweat glandmyoepithelial cells under conditions similar to those in vivo.
This time, we introduced an immortalizing gene into human
eccrine sweat gland myoepithelial cells under culture, which
resulted in the successful generation of iEM cells that can retain
the characters of the human eccrine sweat gland myoepithelial
cells for a long time. Although human eccrine sweat gland
myoepithelial cells could previously be subcultured for onlyaround 3 generations, iEM cells, which our research group hasgenerated, can be subcultured for more than 10 generations. In
addition, iEM cells express α-smooth muscle actin (α-SMA), acharacteristic gene that supports the function of human
myoepithelial cells and which helps maintain the characters ofhuman myoepithelial cells (Fig. 2). Such characteristics are notfound in conventional cultured human eccrine sweat glandcells.

Figure 2.
Viability of immortalized human eccrine sweat
gland myoepithelial (iEM) cells established in thisresearch, and expression of a characteristic gene

Impact of the research achievements on society(The significance of the research achievements)
Our research generated iEM cells that can be cultured for a long time. To control sweating, we need to elucidate theproperties of the cells that make up sweat glands. Advancement of research on human eccrine sweat gland
myoepithelial cells will lead to the development of next-generation antiperspirants that can inhibit sweating byregulating the activity of the cells that make up sweat glands, treatment methods that can restore normal sweating toimprove sweating dysfunction, and techniques for regeneration of sweat glands.

Special notes
Our research achievements, namely iEM cells and the process of their cultivation, are registered as a patent.
Title of Invention: IMMORTALIZED SWEAT GLAND MYOEPITHELIAL CELLSPatent Number: 6563145
Inventors: Tomohisa Hayakawa1, Ryuichiro Kurata1,2, Fumitaka Fujita1,2, Fumihiro Okada1,2, Kiyotoshi Sekiguchi3
1 Laboratory of Advanced Cosmetic Science, Graduate School and School of Pharmaceutical Science, Osaka
University
2 Fundamental Research Institute, Mandom Corporation
3 Department of Contribution Research, Institute for Protein Research, Osaka University
This research was partly supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotionof Science (16K16602: Elucidation of the aging mechanism of sweat glands by cellular senescence analysis, and19K20113: Elucidation of the aging mechanism of sweat glands by immortalized myoepithelial cells).

Glossary※

1. Laboratory of Advanced Cosmetic Science
The Laboratory of Advanced Cosmetic Science is a research laboratory established jointly by the Graduate School
and School of Pharmaceutical Science, Osaka University and Mandom Corporation in June 2015. Joint researchlaboratories receive investments from private companies and other organizations. Researchers from the university
and the investing company conduct research on the same theme on an equal footing, with the aim of achievingbreakthrough research results. The Laboratory of Advanced Cosmetic Science is an independent researchorganization in Osaka University. The university and Mandom Corporation cooperate to conduct research flexibly
and rapidly while exchanging information and creating knowledge.
※
2. Immortalization
Immortalization is a technique that enables extension of the cellular proliferation limit by introducing exogenous
genes that enhance cell proliferation.
※
3. Sweat gland myoepithelial cells
The secreting part of sweat glands (the part that produces sweat) is made up of two types of cells: luminal cells
and myoepithelial cells (Fig. 1). Myoepithelial cells contract the sweat glands to push sweat out onto the bodysurface. They also function as stem cells for sweat glands and therefore play a key role in terms of maintaining
homeostasis of sweat glands.
※
4. Applied Biological Materials Inc.
Applied Biological Materials Inc. is a company dealing with bio-related products, particularly recombinant products.
The company sells recombinant cells and the equipment and products necessary for their production.
#1-3671 Viking Way, Richmond, BC, V6V 2J5, CANADATEL: 604-247-2416
FAX: 604-247-2414
Website: https://www.abmgood.com/about-us.html※5.MTA (Material Transfer Agreement)
The contract between research institutes for the transfer of research materials (loan, transfer, etc.). It stipulates
the handling and attribution of papers and intellectual property rights obtained as a result of research, in addition
to the provisions on the handling of the substance itself.

Contact us<About the research content>
Guest Professor Fumihiro Okada
Laboratory of Advanced Cosmetic Science, Graduate School and School of Pharmaceutical Science, OsakaUniversity
TEL: 06-6105-5792
FAX: 06-6105-5790
E-mail: okada-f@phs.osaka-u.ac.jp
Professor Kiyotoshi Sekiguchi
Department of Contribution Research, Institute for Protein Research, Osaka University
TEL: 06-6105-5935
FAX: 06-6105-5936

E-mail: sekiguch@protein.osaka-u.ac.jp<About this press release>
Public Relations Department Graduate School and School of Pharmaceutical Sciences, Osaka University
TEL：06-6879-8144
FAX: 06-6879-8154
E-mail: yakugaku-syomu@office.osaka-u.ac.jp
Okuda / Kuriyama , Osaka Head Office
Department of Public Relations, Mandom Corporation
TEL: 06-6767-5021
FAX: 06-6767-5045
E-mail: press@mandom.com

Message from the research representative (Hayakawa)
Cultured cells are widely used as a basic experiment method in biological research. However, with recent development ofstem cell research, it has been pointed out that conventional cultured cells differ from those in vivo and are therefore
problematic in terms of a test system. New test systems involving cultured cells that retain their in vivo characteristics bymimicking the natural biostructure have attracted attention as a solution to this problem. Sweat gland cells, unlike skin cells,
originally did not have widely distributed cell lines, and alterations in myoepithelial cells of conventional sweat gland celllines used by some researchers have been reported for the above reason. Our research group focused on the fact that humaneccrine sweat gland myoepithelial cells cultured in an environment similar to that in vivo form a spherical structure covered
with myoepithelial cells. Direct introduction of an immortalizing gene into the spherical structure enabled us to efficiently
immortalize human eccrine sweat gland myoepithelial cells. The iEM cells generated by this method retain the charactersof myoepithelial cells. Therefore, we expect that iEM cells, as a new test system for sweat glands, will help in thedevelopment of antiperspirants, treatment of sweat gland disorders, and regeneration of sweat glands.