Novel phototoxicity assay using human embryonic stem cell-derived retinal pigment epithelial cells
| Autor: | Koichi Saito, Noriyuki Suzuki, Tokushige Nakano, Kiyoshi Higashi, Atsushi Kuwahara, Satoshi Ando, Takashi Mori |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Neutral red Ultraviolet Rays Human Embryonic Stem Cells Gene Expression Retinal Pigment Epithelium Biology Toxicology 030226 pharmacology & pharmacy 3T3 cells Melanin Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine medicine Animals Humans Gene Cells Cultured Melanins Tight junction Cell Differentiation Epithelial Cells Retinal 3T3 Cells Anatomy Embryonic stem cell Coculture Techniques Cell biology 030104 developmental biology medicine.anatomical_structure chemistry Biological Assay sense organs Phototoxicity Dermatitis Phototoxic |
| Zdroj: | Toxicology. 378:1-9 |
| ISSN: | 0300-483X |
| DOI: | 10.1016/j.tox.2017.01.003 |
| Popis: | Some chemicals are harmful in to light-exposed tissues such as skin and eyes. The 3T3 Neutral Red Uptake Phototoxicity Test has been validated and adopted by the Organization of Economic and Community Development (OECD) as a method of evaluating chemical phototoxicity using mouse 3T3 fibroblasts. However, the high rate of false positive results associated with this test eventually led to increased laboratory animal usage. Although the eye is vulnerable to light damage because of constant exposure to environmental radiation, few approaches are available to predict ocular phototoxicity in humans. Here, we propose a tier one test that identifies the potential ocular phototoxicity of chemical substances. Using a three-dimensional culture technique, human embryonic stem cells (hESCs) were differentiated to retinal pigment epithelial cell (RPE) precursors. The precursors after prolonged treatment with FBS formed a uniform hexagonal lattice of cells with well-developed tight junctions and time-dependent elevation of melanin content and RPE maturation marker levels. Hierarchical clustering of gene transcripts revealed that hESC-derived RPEs were very similar to tissue-derived adult RPEs. Interestingly, there were a high percentage of chemicals eliciting a positive response in 3T3 cells and negative in hESC-derived RPEs under the experimental conditions used in the phototoxicity test. The response to treatment of hESC-derived RPEs with these negative chemicals became positive at a higher dose of UVA irradiation; however, the biological responses to these chemicals differed between the two cells. Taken together, we conclude that hESC-derived RPEs are novel tool for future toxicological and mechanistic studies of ocular phototoxicity in humans. |
| Databáze: | OpenAIRE |
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