Evaluation of Retinal Pigment Epithelial Cell Cytotoxicity of Recombinant Tissue Plasminogen Activator Using Human-Induced Pluripotent Stem Cells. (19th March 2019)

• Record Type: Journal Article
• Authors: Kamao, Hiroyuki; Miki, Atsushi; Kiryu, Junichi
• Other Names: Morse Lawrence S. Academic Editor.
• Is Part Of: Journal of ophthalmology. Volume 2019(2019)
• Publication Date: 2019-03-19
• Subjects: Ophthalmology -- Periodicals; Eye Diseases; Ophthalmology; Ophthalmology; Electronic journals; Periodicals; Periodicals; Fulltext; Internet Resources; Periodicals; 617.7
• DOI: 10.1155/2019/7189241
• Languages: English
• ISSNs: 2090-004X

Abstract

Abstract : Purpose . The evaluation of drug-induced cytotoxicity is of great importance for the clinical application of pharmaceutical products, and human-induced pluripotent stem cells (hiPSCs) have received considerable scrutiny as a cell source for in vitro cytotoxicity testing. The aim of this study is to validate the concept of cytotoxicity testing using hiPSC-derived retinal pigment epithelium (hiPSC-RPE) by comparing the responsiveness of human fetal RPE (hfRPE) and human RPE cell line (ARPE19) to recombinant tissue plasminogen activator (rtPA). Methods . HfRPE, two types of hiPSC-RPE, and ARPE19 were cultured in media with or without rtPA. A lactate dehydrogenase release assay was performed to investigate the dose- and time-dependent effects of rtPA on cell death. RPE function was evaluated by measuring the secretion of pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) and RPE-specific gene expression. Results . Rates of cell damage in hfRPE and both hiPS-RPE were increased by rtPA supplementation (2000 and 4000 μ g/ml) for 1 hour, whereas ARPE19 cell damage was increased by supplementation with rtPA at concentrations higher than 50 μ g/ml. Although 100 μ g/ml rtPA for 24 hours did not affect RPE cell function, sustained rtPA exposure induced prolonged cytotoxic effects in hfRPE and two hiPSC-RPE, but not ARPE19. Conclusion . The responsiveness of hiPSC-RPE to rtPA is similar to that of hfRPE in terms of cell death and cell function. Thus, hiPSC-RPE is a valuable cell source for in vitro cytotoxicity testing.