ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells

Abstract 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 c...

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Autores principales:	Yosuke Ida, Fumihito Hikage, Hiroshi Ohguro
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ee4c5cd487c748d1ad04cb06831c534f
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spelling	oai:doaj.org-article:ee4c5cd487c748d1ad04cb06831c534f2021-12-02T11:37:19ZROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells10.1038/s41598-021-84955-72045-2322https://doaj.org/article/ee4c5cd487c748d1ad04cb06831c534f2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-84955-7https://doaj.org/toc/2045-2322Abstract 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.Yosuke IdaFumihito HikageHiroshi OhguroNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yosuke Ida Fumihito Hikage Hiroshi Ohguro ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells
description	Abstract 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	article
author	Yosuke Ida Fumihito Hikage Hiroshi Ohguro
author_facet	Yosuke Ida Fumihito Hikage Hiroshi Ohguro
author_sort	Yosuke Ida
title	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_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_sort	rock inhibitors enhance the production of large lipid-enriched 3d organoids of 3t3-l1 cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/ee4c5cd487c748d1ad04cb06831c534f
work_keys_str_mv	AT yosukeida rockinhibitorsenhancetheproductionoflargelipidenriched3dorganoidsof3t3l1cells AT fumihitohikage rockinhibitorsenhancetheproductionoflargelipidenriched3dorganoidsof3t3l1cells AT hiroshiohguro rockinhibitorsenhancetheproductionoflargelipidenriched3dorganoidsof3t3l1cells
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ROCK inhibitors enhance the production of large lipid-enriched 3D organoids of 3T3-L1 cells

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