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Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation

Background: Three-dimensional (3D) culture changes cell characteristics and function, suggesting that 3D culture provides a more physiologically relevant environment for cells compared with 2D culture. We investigated the differences in cell functions depending on the culture model in human trophobl...

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Autores principales: Seno, Kotomi, Munakata, Yasuhisa, Sano, Michiya, Kawahara-Miki, Ryouka, Takahashi, Hironori, Ohkuchi, Akihide, Iwata, Hisataka, Kuwayama, Takehito, Shirasuna, Koumei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6121648/
https://www.ncbi.nlm.nih.gov/pubmed/30096769
http://dx.doi.org/10.3390/ijms19082322
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author Seno, Kotomi
Munakata, Yasuhisa
Sano, Michiya
Kawahara-Miki, Ryouka
Takahashi, Hironori
Ohkuchi, Akihide
Iwata, Hisataka
Kuwayama, Takehito
Shirasuna, Koumei
author_facet Seno, Kotomi
Munakata, Yasuhisa
Sano, Michiya
Kawahara-Miki, Ryouka
Takahashi, Hironori
Ohkuchi, Akihide
Iwata, Hisataka
Kuwayama, Takehito
Shirasuna, Koumei
author_sort Seno, Kotomi
collection PubMed
description Background: Three-dimensional (3D) culture changes cell characteristics and function, suggesting that 3D culture provides a more physiologically relevant environment for cells compared with 2D culture. We investigated the differences in cell functions depending on the culture model in human trophoblast cells (Sw.71). Methods: Sw.71 cells were incubated in 2D monolayers or simple 3D spheroids. After incubation, cells were corrected to assess RNA-seq transcriptome or protein expression, and culture medium were corrected to detect cytokines. To clarify the role of actin cytoskeleton, spheroid Sw.71 cells were treated mycalolide B (inhibitor of actin polymerization) in a 3D culture. Results: RNA-seq transcriptome analysis, results revealed that 3D-cultured cells had a different transcriptional profile compared with 2D-cultured cells, especially regarding inflammation-related molecules. Although interleukin-6 (IL-6) mRNA level was higher in 3D-culured cells, its secretion levels were higher in 2D-cultured cells. In addition, the levels of mRNA and protein expression of regnase-1, regulatory RNase of inflammatory cytokine, significantly increased in 3D culture, suggesting post-translational modification of IL-6 mRNA via regnase-1. Treatment with mycalolide B reduced cell-to-cell contact to build 3D formation and increased expression of actin cytoskeleton, resulting in increased IL-6 secretin. Conclusion: Cell dimensionality plays an essential role in governing the spatiotemporal cellular outcomes, including inflammatory cytokine production and its negative regulation associated with regnase-1.
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spelling pubmed-61216482018-09-07 Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation Seno, Kotomi Munakata, Yasuhisa Sano, Michiya Kawahara-Miki, Ryouka Takahashi, Hironori Ohkuchi, Akihide Iwata, Hisataka Kuwayama, Takehito Shirasuna, Koumei Int J Mol Sci Article Background: Three-dimensional (3D) culture changes cell characteristics and function, suggesting that 3D culture provides a more physiologically relevant environment for cells compared with 2D culture. We investigated the differences in cell functions depending on the culture model in human trophoblast cells (Sw.71). Methods: Sw.71 cells were incubated in 2D monolayers or simple 3D spheroids. After incubation, cells were corrected to assess RNA-seq transcriptome or protein expression, and culture medium were corrected to detect cytokines. To clarify the role of actin cytoskeleton, spheroid Sw.71 cells were treated mycalolide B (inhibitor of actin polymerization) in a 3D culture. Results: RNA-seq transcriptome analysis, results revealed that 3D-cultured cells had a different transcriptional profile compared with 2D-cultured cells, especially regarding inflammation-related molecules. Although interleukin-6 (IL-6) mRNA level was higher in 3D-culured cells, its secretion levels were higher in 2D-cultured cells. In addition, the levels of mRNA and protein expression of regnase-1, regulatory RNase of inflammatory cytokine, significantly increased in 3D culture, suggesting post-translational modification of IL-6 mRNA via regnase-1. Treatment with mycalolide B reduced cell-to-cell contact to build 3D formation and increased expression of actin cytoskeleton, resulting in increased IL-6 secretin. Conclusion: Cell dimensionality plays an essential role in governing the spatiotemporal cellular outcomes, including inflammatory cytokine production and its negative regulation associated with regnase-1. MDPI 2018-08-08 /pmc/articles/PMC6121648/ /pubmed/30096769 http://dx.doi.org/10.3390/ijms19082322 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Seno, Kotomi
Munakata, Yasuhisa
Sano, Michiya
Kawahara-Miki, Ryouka
Takahashi, Hironori
Ohkuchi, Akihide
Iwata, Hisataka
Kuwayama, Takehito
Shirasuna, Koumei
Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation
title Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation
title_full Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation
title_fullStr Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation
title_full_unstemmed Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation
title_short Aggregation of Human Trophoblast Cells into Three-Dimensional Culture System Enhances Anti-Inflammatory Characteristics through Cytoskeleton Regulation
title_sort aggregation of human trophoblast cells into three-dimensional culture system enhances anti-inflammatory characteristics through cytoskeleton regulation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6121648/
https://www.ncbi.nlm.nih.gov/pubmed/30096769
http://dx.doi.org/10.3390/ijms19082322
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