In vitro formation of the Merkel cell‐neurite complex in embryonic mouse whiskers using organotypic co‐cultures

Ishida, Kentaro; Saito, Tetsuichiro; Mitsui, Toshiyuki

doi:10.1111/dgd.12535

Cited by 3 publications

(3 citation statements)

References 38 publications

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“…Fluorescent immunohistochemistry using 8‐μm fresh frozen sections were performed as previously described (Ishida, Saito, & Mitsui, ). Sections were incubated with the following primary antibodies: anti‐cleaved caspase‐3 antibody (5A1E; 1:100; rabbit; CST), anti‐β‐catenin antibody (1:1000; rabbit; Sigma), anti‐ZO‐1 antibody (ZO1‐1A12; 1:800; mouse; Life Technologies) and phospho‐Histone H3 (Ser10) antibody (1:100; mouse; CST).…”

Section: Methodsmentioning

confidence: 99%

Involvement of selective epithelial cell death in the formation of feather buds on a bioengineered skin

2019

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Selective cell death by apoptosis plays important roles in organogenesis. Apoptotic cells are observed in the developmental and homeostatic processes of several ectodermal organs, such as hairs, feathers, and mammary glands. In chick feather development, apoptotic events have been observed during feather morphogenesis, but have not been investigated during early feather bud formation. Previously, we have reported a method for generating feather buds on a bioengineered skin from dissociated skin epithelial and mesenchymal cells in three‐dimensional culture. During the development of the bioengineered skin, epithelial cavity formation by apoptosis was observed in the epithelial tissue. In this study, we examined the selective epithelial cell death during the bioengineered skin development. Histological analyses suggest that the selective epithelial cell death in the bioengineered skin was induced by caspase‐3‐related apoptosis. The formation of feather buds of the bioengineered skin was disturbed by the treatment with a pan‐caspase inhibitor. The pan‐caspase inhibitor treatment suppressed the rearrangement of the epithelial layer and the formation of dermal condensation, which are thought to be essential step to form feather buds. The suppression of the formation of feather buds on the pan‐caspase inhibitor‐treated skin was partially compensated by the addition of a GSK‐3β inhibitor, which activates Wnt/β‐catenin signaling. These results suggest that the epithelial cell death is involved in the formation of feather buds of the bioengineered skin. These observations also suggest that caspase activities and Wnt/β‐catenin signaling may contribute to the formation of epithelial and mesenchymal components in the bioengineered skin.

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Section: Methodsmentioning

confidence: 99%

Involvement of selective epithelial cell death in the formation of feather buds on a bioengineered skin

2019

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“…The cellular process of co-culture can be extended to other cell lines to further investigate cell-to-cell interaction. This method was developed by Ishida et al [27]. Two methods of culturing mustache pads and trigeminal ganglion explants were combined in a co-culture model.…”

Section: Skin-nerves Interplay In Skin Homeostasismentioning

confidence: 99%

Emergence of New Concepts in Skin Physiopathology through the Use of in vitro Human Skin Explants Models

Cousin,

Misery,

de Vries

et al. 2023

Dermatology

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Background: This review summarizes uses and new applications for dermatological research of in vitro culture models of human skin explants (HSEs). In the last decade, many innovations have appeared in the literature and an exponential number of studies have been recorded in various fields of application such as process culture engineering, stem cell extractions methodology, or cell-to-cell interaction studies under physiological and pathological conditions, wound-healing, and inflammation. Most studies also concerned pharmacology, cosmetology, and photobiology. However, these topics will not be considered in our review. Summary: A better understanding of the mechanisms driving intercellular relationships, at work in the maintenance of 3D tissue architectures has led to the improvement of cell culture techniques. Many papers have focused on the physiological ways that govern in vitro tissue maintenance of HSEs. The analysis of the necessary mechanical stress, intercellular and cell-matrix interactions, allows the maintenance and prolonged use of HSEs in culture for up to 15 days, regardless of the great variability of study protocols from one laboratory to another and in accordance with the objectives set. Because of their close similarities to fresh skin, HSEs are increasingly used to study skin barrier repair and wound healing physiology. Easy to use in co-culture, this model allows a better understanding of the connections and interactions between the peripheral nervous system, the skin and the immune system. The development of the concept of an integrated neuro-immuno-cutaneous system at work in skin physiology and pathology highlighted by this article represents one of the new technical challenges in the field of in vitro culture of HSE. This review of the literature also reveals the importance of using such models in pathology. As sources of stem cells, HSEs are the basis for the development of new tissue engineering models such as organoids or optical clearing tissues technology. This study identifies the main advances and cross-cutting issues in the use of HSE.

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“…To study the formation of MC complexes in the whiskers, an organotypic co-culture model was developed [ 137 ]. The dynamic process of formation of MC complexes cannot be observed directly due to embryonic development.…”

Section: In Vitro Studies (Culture Of Merkel Cells)mentioning

confidence: 99%

Merkel Cells Are Multimodal Sensory Cells: A Review of Study Methods

Bataille

Gall

Miséry

et al. 2022

Cells

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Merkel cells (MCs) are rare multimodal epidermal sensory cells. Due to their interactions with slowly adapting type 1 (SA1) Aβ low-threshold mechanoreceptor (Aβ-LTMRs) afferents neurons to form Merkel complexes, they are considered to be part of the main tactile terminal organ involved in the light touch sensation. This function has been explored over time by ex vivo, in vivo, in vitro, and in silico approaches. Ex vivo studies have made it possible to characterize the topography, morphology, and cellular environment of these cells. The interactions of MCs with surrounding cells continue to be studied by ex vivo but also in vitro approaches. Indeed, in vitro models have improved the understanding of communication of MCs with other cells present in the skin at the cellular and molecular levels. As for in vivo methods, the sensory role of MC complexes can be demonstrated by observing physiological or pathological behavior after genetic modification in mouse models. In silico models are emerging and aim to elucidate the sensory coding mechanisms of these complexes. The different methods to study MC complexes presented in this review may allow the investigation of their involvement in other physiological and pathophysiological mechanisms, despite the difficulties in exploring these cells, in particular due to their rarity.

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In vitro formation of the Merkel cell‐neurite complex in embryonic mouse whiskers using organotypic co‐cultures

Cited by 3 publications

References 38 publications

Involvement of selective epithelial cell death in the formation of feather buds on a bioengineered skin

Involvement of selective epithelial cell death in the formation of feather buds on a bioengineered skin

Emergence of New Concepts in Skin Physiopathology through the Use of in vitro Human Skin Explants Models

Merkel Cells Are Multimodal Sensory Cells: A Review of Study Methods

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