October 27, 2016

In Pursuit of a Next-generation Antiperspirant
Mandom Succeeds in Isolation of Sweat Gland Stem Cells Possessing
the Ability to Regenerate Sweat Gland-like Structures In Vitro
Mandom Corporation (Headquarters: Osaka; President Executive Officer: Motonobu Nishimura; hereafter,Mandom) has, as part of its Laboratory of Advanced Cosmetic Science (Collaborative Research Laboratory between
Osaka University Graduate School of Pharmaceutical Sciences and Mandom), been involved in the development offoundational technologies for the creation of next-generation deodorants for the control of sweating.
At present, in collaboration with Osaka University’s Institute for Protein Research, Graduate School of Medicine,and Graduate School of Pharmaceutical Sciences, we investigated the replication of sweating function.
Previous studies have revealed that stem cells are present in epidermis and hair follicles of human; they contribute to
the maintenance of the functions of each organ. However, it has remain unknown whether existence of stem cells inhuman sweat glands, because the procedure for isolation of sweat gland cells have not been established.
Therefore, we first focused on a method for isolating sweat gland cells from the human skin (with the approval ofthe ethics committee) and identified the markers that accurately discriminate the compartments consisting of humansweat glands. In addition, we identified sweat gland stem cells within sweat gland cells and succeeded in regenerating asweat gland-like structure in vitro from the sweat gland stem cells. These results led to improved methods for theevaluation of the sweating function of the sweat glands and enabled the direct application of antiperspirants—medicines
that, primarily, seal off sweat glands—to sweat glands. This enabled the proposition of a new mechanism of action for
antiperspirants—one that “improves the amount and quality of sweating.”
We plan to present these findings at the International Federation of Societies of Cosmetic Chemists’ Orlando
Conference, held in the United States between October 30 and November 2, 2016.
1. Establishment of a Method for Isolating Sweat Glands from the Human Skin Tissue
The mechanism of sweating from sweat glands has not yet been investigated. In recent years, remarkable progresshas been made in the application of regenerative medicine technologies to replicate sweat glands as a techniqueapplicable to the treatment of diseases specific to the sweat glands. To resolve unpleasant sudomotor dysfunction such as
excessive sweating or smell, we focused on the improvement of sweating function by regenerating sweat glands. To
clarify the localization of stem cells in the sweat glands, we first developed a method for the high-purity isolation of fourcell populations consisting of human sweat glands.
Human sweat glands divided into the duct that carries sweat to the skin surface and the secretory portion that
produces and releases sweat. The duct is composed of luminal cells and basal cells, and the secretory portion iscomposed of luminal cells and myoepithelial cells (Figure 1). We identified the markers (αSMA, Keratin8, S100P, andS100A2) that exclusively distinguish four populations consisting of sweat gland (Figure 2).

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-1-This enabled the establishment of a method for isolating highly pure luminal cells and myoepithelial cells from thesecretory portions of the human sweat glands (Figure 3). This method not only allowed the analysis of sweat gland stem
cells, but also can be applied to various kinds of sweat gland studies

2. Discovery of Sweat Gland Stem Cells that Maintain Sweat Gland Function
Using the established sweat gland cell isolation method, we sought to investigate the population that contain sweat
gland stem cells. To demonstrate the sweat gland stem cells, we have to substantiate whether the cells possess selfrenewal ability (ability to replicate on its own multiple times) and multipotency (ability to change into multiple celltypes). We evaluated whether the sweat gland cell compartments isolated from the human skin tissues possess selfrenewal ability and multipotency and found that myoepithelial cells possess these abilities. Thus, we clarified that themyoepithelial cells are sweat gland stem cells (Figure 4).
3. Success in the Regeneration of Sweat Gland-like Structures from Sweat Gland Stem Cells
Success in the Regeneration of Sweat Gland-like Structures from Sweat Gland Stem Cells
After using αSMA and Keratin8 to assess the cross-sections of the spheres generated from myoepithelial cells, wefound that αSMA, a marker for myoepithelial cells, was detected on the outer layers of the sphere, whereas Keratin8, amarker for luminal cells, was detected in the inner part of the sphere, indicating that the outer and inner parts of thespheres were composed of myoepithelial cells and luminal cells, respectively. These findings revealed that the sphereregenerated from myoepithelial cells is a sweat gland-like structure (Figure 5).
In the future, we intend to investigate the sweating function of sweat glands using the sweat gland-like structure.
Mandom will continue to proactively prioritize the effects and feelings of consumers in the development of highly
effective antiperspirants that can satisfy those that use them.
[Reference Material]
Figure 1: The Four Cell Populations that Consist of the Human Sweat Gland
Duct
Luminalcells

Epidermis

Stem cells

Secretory
portionHair follicle
Sweat gland

Basal cells
Luminal cells

Myoepithelial
cells-2-Figure 2: Identifying Markers that Exclusively Distinguish Sweat Gland Components
Exclusively Distinguish Sweat Gland Components

cross-sectional viewDuct(Duct, luminal cell)(Secretory gland, luminalcell)(Duct, luminal cell)(Secretory gland, myoepithelialcell)(Duct, basal cell)(Secretory gland, luminalcell)(Duct, basal cell)(Secretory gland, myoepithelialcell)

Luminalcells

Luminal cells

Secretory
portion Basal cells

Myoepithelialcells

Figure 3: Isolation of Myoepithelial Cells and Luminal Cells from a Secretory Gland by using αSMA
and Keratin8
Secretory gland cells
Luminal cells

Percentage of positive cells(Luminal cell marker)(Myoepithelial cell marker)

Myoepithelial cells

Myoepithelial cellsLuminal cells

Figure 4: Stem Cell Characteristics of Myoepithelial Cells (Pluripotency and Self-replication)
Myoepithelial cells

Cell mass
Pluripotency

Stem cells-3-

Self-replication

Figure 5: Sweat Gland-like Structure Replicated from the Sweat Gland Stem Cells
Sweat gland-like structure

Sweat gland cells

Sweat gland

Cell mass(Myoepithelial cells)(Luminal cells)-4-