TGF-β effects on adipogenesis of 3T3-L1 cells differ in 2D and 3D cell culture conditions.

Autor: Umetsu A; Department of Ophthalmology, Sapporo Medical University School of Medicine, Japan., Watanabe M; Department of Ophthalmology, Sapporo Medical University School of Medicine, Japan., Sato T; Department of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Japan.; Department of Cellular Physiology and Signal Transduction, Sapporo Medical University School of Medicine, Japan., Higashide M; Department of Ophthalmology, Sapporo Medical University School of Medicine, Japan., Nishikiori N; Department of Ophthalmology, Sapporo Medical University School of Medicine, Japan., Furuhashi M; Department of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Japan., Ohguro H; Department of Ophthalmology, Sapporo Medical University School of Medicine, Japan.
Jazyk: angličtina
Zdroj: FEBS open bio [FEBS Open Bio] 2024 Dec; Vol. 14 (12), pp. 2026-2037. Date of Electronic Publication: 2024 Oct 08.
DOI: 10.1002/2211-5463.13890
Abstrakt: The TGF-β superfamily plays a pivotal role in the regulation of adipogenesis, but little is known about the potential differential role of the three isoforms of TGF-β, TGF-β-1~3. To further elucidate their role, two-dimensionally (2D) and three-dimensionally (3D) cultured 3T3-L1 mouse preadipocytes were subjected to the following analyses: (a) qPCR analysis of adipogenesis-related factors and major extracellular matrix protein (2D and /or 3D), (b) lipid staining by Oil Red O (2D) or BODIPY (3D), (c) Seahorse cellular metabolic measurement (2D), and (d) size and stiffness measurements of 3D 3T3-L1 spheroids. In the 2D cultured 3T3-L1 cells, mRNA expression levels of adipogenesis-related genes and Oil Red O lipid staining intensity were significantly increased by adipogenesis and they were substantially decreased following treatment with 0.1 nm TGF-β isoforms, with TGF-β2 having the greater effects. Consistent with these results, treatment with TGF-β2 resulted in suppression of mitochondrial and glycolytic functions in 2D cultured 3T3-L1 cells. However, the inhibitory effect of TGF-β on adipogenesis decreased under 3D spheroid culture conditions and TGF-β isoforms did not affect adipogenesis-induced (a) enlargement and downsizing of 3T3-L1 spheroids, (b) increase in BODIPY lipid staining intensity, and (c) up-regulation of the mRNA expression of adipogenesis-related genes. The findings presented herein suggest that the three TGF-β isoforms have different suppressive effects on adipogenesis-related cellular properties of 2D cultured 3T3-L1 cells and that their effects decrease under 3D spheroid culture conditions.
(© 2024 The Author(s). FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)
Databáze: MEDLINE
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