Experimental Cell Research

2015

DOI: 10.1016/j.yexcr.2015.01.014

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Matrigel basement membrane matrix induces eccrine sweat gland cells to reconstitute sweat gland-like structures in nude mice

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Cited by 28 publications

(19 citation statements)

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“…In an in vitro 3D model, orientation and positioning is controlled by the apical-basal axis, which determines the tubular tissue architecture [ 35 ]. The responsible relationships for differentiation of epithelia during 3D reconstruction like the interaction of cells with extracellular matrix (ECM) and the surrounding media [ 36 ] could indicate these regulatory processes leading to apical-basal tissue architecture in our 3D SG model. Further research is required to provide evidence of matrix existence and of distinct tissue types.…”

Section: Resultsmentioning

confidence: 99%

A novel organotypic 3D sweat gland model with physiological functionality

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et al. 2017

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Dysregulated human eccrine sweat glands can negatively impact the quality-of-life of people suffering from disorders like hyperhidrosis. Inability of sweating can even result in serious health effects in humans affected by anhidrosis. The underlying mechanisms must be elucidated and a reliable in vitro test system for drug screening must be developed. Here we describe a novel organotypic three-dimensional (3D) sweat gland model made of primary human eccrine sweat gland cells. Initial experiments revealed that eccrine sweat gland cells in a two-dimensional (2D) culture lose typical physiological markers. To resemble the in vivo situation as close as possible, we applied the hanging drop cultivation technology regaining most of the markers when cultured in its natural spherical environment. To compare the organotypic 3D sweat gland model versus human sweat glands in vivo, we compared markers relevant for the eccrine sweat gland using transcriptomic and proteomic analysis. Comparing the marker profile, a high in vitro-in vivo correlation was shown. Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), muscarinic acetylcholine receptor M3 (CHRM3), Na+-K+-Cl- cotransporter 1 (NKCC1), calcium-activated chloride channel anoctamin-1 (ANO1/TMEM16A), and aquaporin-5 (AQP5) are found at significant expression levels in the 3D model. Moreover, cholinergic stimulation with acetylcholine or pilocarpine leads to calcium influx monitored in a calcium flux assay. Cholinergic stimulation cannot be achieved with the sweat gland cell line NCL-SG3 used as a sweat gland model system. Our results show clear benefits of the organotypic 3D sweat gland model versus 2D cultures in terms of the expression of essential eccrine sweat gland key regulators and in the physiological response to stimulation. Taken together, this novel organotypic 3D sweat gland model shows a good in vitro-in vivo correlation and is an appropriate alternative for screening of potential bioactives regulating the sweat mechanism.

“…In an in vitro 3D model, orientation and positioning is controlled by the apical-basal axis, which determines the tubular tissue architecture [ 35 ]. The responsible relationships for differentiation of epithelia during 3D reconstruction like the interaction of cells with extracellular matrix (ECM) and the surrounding media [ 36 ] could indicate these regulatory processes leading to apical-basal tissue architecture in our 3D SG model. Further research is required to provide evidence of matrix existence and of distinct tissue types.…”

Section: Resultsmentioning

confidence: 99%

A novel organotypic 3D sweat gland model with physiological functionality

¹

,

²

,

³

et al. 2017

View full text Add to dashboard Cite

Dysregulated human eccrine sweat glands can negatively impact the quality-of-life of people suffering from disorders like hyperhidrosis. Inability of sweating can even result in serious health effects in humans affected by anhidrosis. The underlying mechanisms must be elucidated and a reliable in vitro test system for drug screening must be developed. Here we describe a novel organotypic three-dimensional (3D) sweat gland model made of primary human eccrine sweat gland cells. Initial experiments revealed that eccrine sweat gland cells in a two-dimensional (2D) culture lose typical physiological markers. To resemble the in vivo situation as close as possible, we applied the hanging drop cultivation technology regaining most of the markers when cultured in its natural spherical environment. To compare the organotypic 3D sweat gland model versus human sweat glands in vivo, we compared markers relevant for the eccrine sweat gland using transcriptomic and proteomic analysis. Comparing the marker profile, a high in vitro-in vivo correlation was shown. Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), muscarinic acetylcholine receptor M3 (CHRM3), Na+-K+-Cl- cotransporter 1 (NKCC1), calcium-activated chloride channel anoctamin-1 (ANO1/TMEM16A), and aquaporin-5 (AQP5) are found at significant expression levels in the 3D model. Moreover, cholinergic stimulation with acetylcholine or pilocarpine leads to calcium influx monitored in a calcium flux assay. Cholinergic stimulation cannot be achieved with the sweat gland cell line NCL-SG3 used as a sweat gland model system. Our results show clear benefits of the organotypic 3D sweat gland model versus 2D cultures in terms of the expression of essential eccrine sweat gland key regulators and in the physiological response to stimulation. Taken together, this novel organotypic 3D sweat gland model shows a good in vitro-in vivo correlation and is an appropriate alternative for screening of potential bioactives regulating the sweat mechanism.

“…The discovery that reconstituted cells from human eccrine glands could form sweat gland‐like structures in culture is also an interesting development in the quest to assist burn victims. However, this technique, too, needs further investigation.…”

Section: Regnerating Sweat Glandsmentioning

confidence: 99%

The evolution of eccrine sweat gland research towards developing a model for human sweat gland function

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2018

Experimental Dermatology

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For several decades now, researchers, professional bodies, governments, and journals such as the journal of Experimental Dermatology have worked to reduce the number of animals used in experimentation. This review centres on investigations into how human sweat glands produce sweat and how that research has evolved over the years. It is hoped that this review will show that as methodologies advanced, sweat gland research has come to rely less and less on a variety of animal models as investigative tools and information is being primarily obtained through human and mouse material, with a view to further reductions in using animal models.

“…In a previous study, we have established an in vivo 3D reconstitution model of eccrine sweat glands by subcutaneously implanting Matrigel-embedded eccrine sweat gland cells into the subcutis of nude mice (Li et al 2015a). In the present study, we have examined the changes of keratins and alpha-SMA at various developmental time points in order to investigate the development and cell differentiation of eccrine sweat gland cells during the course of 3D reconstitution.…”

Section: Introductionmentioning

confidence: 99%

Changes in keratins and alpha-smooth muscle actin during three-dimensional reconstitution of eccrine sweat glands

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et al. 2016

Cell Tissue Res

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We have examined the changes of keratins and alpha-SMA at various time points in order to investigate the development and differentiation of eccrine sweat gland cells during the course of three-dimensional (3D) reconstitution. Mixtures of eccrine sweat gland cells and Matrigel were injected subcutaneously into the inguinal regions of nude mice. At 1, 2, 4, 6, 8, 14, 21, 28, 35, and 42 days post-implantation, Matrigel plugs were removed and immunostained. We found that during 3D reconstitution, keratin and alpha-SMA expression changed in a time-dependent manner. At day 1, all cells stained positively for keratin isoforms K5, K14, and K15, with the staining intensity of K15 being weak and K5 and K14 being strong, but none of the cells displayed K7, K8, or alpha-SMA. As time progressed, spheroid-like structures formed with the inner layer acquiring K7 and K8, but losing K5 and K14 expression, and the outer layer acquiring alpha-SMA expression, but losing K15 expression. K8 expression was first noted at day 14, and K7 and alpha-SMA at day 21. The loss of K15 expression was first noted at day 14, K14 at day 21, and K5 at day 28. At 28, 35, and 42 days, the spheroid-like structures could be distinguished, by immunohistochemistry, as having secretory coil-like and coiled duct-like structures. We conclude that the changes in expression of keratins and alpha-SMA in 3D-reconstituted eccrine sweat glands are similar to those of native eccrine sweat glands, indicating that the 3D reconstitution of sweat glands provides an excellent model for studying the development, cytodifferentiation, and regulation of eccrine sweat glands.

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