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ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells
Since the recent discovery of prostaglandin-associated peri-orbitopathy, a great deal of interest has developed concerning the side effects of anti-glaucoma medications toward periocular fatty tissue, especially their adipogenesis. Two- or three-dimension (2D or 3D) cultures of the 3T3-L1 cells were...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7943807/ https://www.ncbi.nlm.nih.gov/pubmed/33750898 http://dx.doi.org/10.1038/s41598-021-84955-7 |
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author | Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi |
author_facet | Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi |
author_sort | Ida, Yosuke |
collection | PubMed |
description | Since the recent discovery of prostaglandin-associated peri-orbitopathy, a great deal of interest has developed concerning the side effects of anti-glaucoma medications toward periocular fatty tissue, especially their adipogenesis. Two- or three-dimension (2D or 3D) cultures of the 3T3-L1 cells were employed to elucidate the effects of the Rho-associated coiled-coil containing protein kinase inhibitor (ROCK-i) the anti-glaucoma drug, Ripasudil, and other ROCK-i, such as Y27632 on adipogenesis. Ultrastructure by electron microscopy and physical stiffness measurements by a micro-squeezer demonstrated the 3D organoids had essentially matured during the 7-day culture. The effects of ROCK-i on 3D organoid sizes, lipid staining, the mRNA expression of adipogenesis related genes, Pparγ, Cebpa and Leptin, and extracellular matrix (ECM) including collagen (COL) 1, 4 and 6, and fibronectin, and physical stiffness were then conducted. Upon adipogenesis, the sizes, lipid staining and mRNA expressions of adipogenesis related genes, Col 4 and Col 6 were dramatically increased, and were further enhanced by ROCK-i. Micro-squeezer analysis demonstrated that adipogenesis resulted in a marked less stiffed 3D organoid and this was further enhanced by ROCK-i. Our present study indicates that ROCK-i significantly enhanced the production of large lipid-enriched 3T3-L1 3D organoids. |
format | Online Article Text |
id | pubmed-7943807 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-79438072021-03-10 ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi Sci Rep Article Since the recent discovery of prostaglandin-associated peri-orbitopathy, a great deal of interest has developed concerning the side effects of anti-glaucoma medications toward periocular fatty tissue, especially their adipogenesis. Two- or three-dimension (2D or 3D) cultures of the 3T3-L1 cells were employed to elucidate the effects of the Rho-associated coiled-coil containing protein kinase inhibitor (ROCK-i) the anti-glaucoma drug, Ripasudil, and other ROCK-i, such as Y27632 on adipogenesis. Ultrastructure by electron microscopy and physical stiffness measurements by a micro-squeezer demonstrated the 3D organoids had essentially matured during the 7-day culture. The effects of ROCK-i on 3D organoid sizes, lipid staining, the mRNA expression of adipogenesis related genes, Pparγ, Cebpa and Leptin, and extracellular matrix (ECM) including collagen (COL) 1, 4 and 6, and fibronectin, and physical stiffness were then conducted. Upon adipogenesis, the sizes, lipid staining and mRNA expressions of adipogenesis related genes, Col 4 and Col 6 were dramatically increased, and were further enhanced by ROCK-i. Micro-squeezer analysis demonstrated that adipogenesis resulted in a marked less stiffed 3D organoid and this was further enhanced by ROCK-i. Our present study indicates that ROCK-i significantly enhanced the production of large lipid-enriched 3T3-L1 3D organoids. Nature Publishing Group UK 2021-03-09 /pmc/articles/PMC7943807/ /pubmed/33750898 http://dx.doi.org/10.1038/s41598-021-84955-7 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells |
title | ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells |
title_full | ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells |
title_fullStr | ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells |
title_full_unstemmed | ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells |
title_short | ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells |
title_sort | rock inhibitors enhance the production of large lipid-enriched 3d organoids of 3t3-l1 cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7943807/ https://www.ncbi.nlm.nih.gov/pubmed/33750898 http://dx.doi.org/10.1038/s41598-021-84955-7 |
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