TGF-β Isoforms and Local Environments Greatly Modulate Biological Nature of Human Retinal Pigment Epithelium Cells

• Published in: Bioengineering (Basel) Vol. 11; no. 6; p. 581
• Main Authors: Nishikiori, Nami, Sato, Tatsuya, Ogawa, Toshifumi, Higashide, Megumi, Umetsu, Araya, Suzuki, Soma, Furuhashi, Masato, Ohguro, Hiroshi, Watanabe, Megumi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 07.06.2024; MDPI
• Subjects: 3D culture; Actin; Biological effects; Cell culture; E-cadherin; Electrical resistivity; Epithelium; Extracellular matrix; Gene expression; Glycolysis; Growth factors; human retinal pigment epithelium; Hypoxia; Hypoxia-inducible factor 1a; Hypoxia-inducible factors; Instrument industry; Isoforms; Oxygen; Pathogenesis; Peroxisome proliferator-activated receptors; Physiology; Pigments; Retina; Retinal pigment epithelium; seahorse cellular metabolic functions; Smooth muscle; Spheroids; Stem cells; TGF-β isoforms; Transforming growth factor-b; Transforming growth factor-b1; Transforming growth factors; Vascular endothelial growth factor; Zonula occludens-1 protein; United States; Japan; St Louis Missouri; United States > US; hypoxia; TGF-β isoforms; 3D culture; human retinal pigment epithelium; seahorse cellular metabolic functions
• ISSN: 2306-5354; 2306-5354
• DOI: 10.3390/bioengineering11060581

To characterize transforming growth factor-β (TGF-β) isoform (TGF-β1~3)-b's biological effects on the human retinal pigment epithelium (RPE) under normoxia and hypoxia conditions, ARPE19 cells cultured by 2D (two-dimensional) and 3D (three-dimensional) conditions were subjected to various analyses, including (1) an analysis of barrier function by trans-epithelial electrical resistance (TEER) measurements; (2) qPCR analysis of major ECM molecules including , , and ; ; ; and , a master regulator for mitochondrial respiration;, tight junction-related molecules, and ; and ; (3) physical property measurements of 3D spheroids; and (4) cellular metabolic analysis. Diverse effects among TGF-β isoforms were observed, and those effects were also different between normoxia and hypoxia conditions: (1) TGF-β1 and TGF-β3 caused a marked increase in TEER values, and TGF-β2 caused a substantial increase in TEER values under normoxia conditions and hypoxia conditions, respectively; (2) the results of qPCR analysis supported data obtained by TEER; (3) 3D spheroid sizes were decreased by TGF-β isoforms, among which TGF-β1 had the most potent effect under both oxygen conditions; (4) 3D spheroid stiffness was increased by TGF-β2 and TGF-β3 or by TGF-β1 and TGF-β3 under normoxia conditions and hypoxia conditions, respectively; and (5) the TGF-β isoform altered mitochondrial and glycolytic functions differently under oxygen conditions and/or culture conditions. These collective findings indicate that the TGF-β-induced biological effects of 2D and 3D cultures of ARPE19 cells were substantially diverse depending on the three TGF-β isoforms and oxygen levels, suggesting that pathological conditions including epithelial-mesenchymal transition (EMT) of the RPE may be exclusively modulated by both factors.