3D CULTURE OF HaCaT KERATINOCYTE CELL LINE AS AN in vitro TOXICITY MODEL

Year 2022, Volume: 23 Issue: 2, 211 - 220, 15.10.2022

Selinay Şenkal , Derya Burukçu Taha Bartu Hayal , Binnur Kıratlı Hatice Burcu Şişli Derya Sağraç , Burçin Asutay , Engin Sümer Fikrettin Şahin , Ayşegül Doğan

https://doi.org/10.23902/trkjnat.1158811

Cited By: 1

Abstract

Ex vivo dermal toxicology analyses are crucial for replacement of in vivo test methods and have been of interest in recent years for testing cosmetics, drugs, and chemicals. Development of an appropriate reconstructed epidermis model might overcome the limitations of monolayer culture systems. In the current study, we used the immortalized human keratinocyte cell line (HaCaT) to develop an ex vivo 3D cell culture system for keratinocyte-based toxicity analysis. Mouse embryonic fibroblast-conditioned medium and Matrigel matrix-based 3D HaCaT cell culture systems expressed skin-related genes and proteins in culture. The 3D HaCaT cultures demonstrated a skin-like phenotype and response against selected test compounds. Reliable results were obtained compared to monolayer HaCaT cells which were exposed to selected chemicals for 1 h and 24 h. Gene expression profiles of 3D HaCaT cell cultures and monolayer cultures were completely different after administration of the test compounds.
Overall, our results showed that a 3D HaCaT cell culture system generated in Matrigel matrix exerted a skin epidermis-like phenotype. Consequently, 3D HaCaT cell cultures may be an acceptable test method for conducting in vitro toxicology experiments. 

Keywords

skin irritation, epidermis, Reconstructed skin, 3D skin model

Thanks

This study was conducted in Yeditepe University. We would like to thank Yeditepe University Writing Center and Hakan Şentürk (İstanbul-TÜRKİYE) for language editing and proofreading support.

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