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hTERT-immortalized Neonatal Melanocytes – an Advanced In Vitro Cell-based Model for Pigmentation and Toxicity Studies

Poster
Green and blue kidney epithelial melanocyte cells.

SOT 61th Annual Meeting and ToxExpo

San Diego, California, United States

March 28, 2022

Abstract

Pigmentation involves two main steps. First, melanocytes perform a myriad of complex biochemical and physiological steps to produce, package, and exocytose melanin containing melanosomes. Secondly, the melanosomes are taken up by neighboring keratinocytes where the stored melanin protects underlying tissues from damaging UV radiation. This process of pigmentation involves a complex interplay between genetic, endocrine, and environmental factors. Primary cells offer one model system to study pigmentation and dermal agents that may disrupt the melanogenesis; however, they are hindered somewhat by donor-to-donor variability and limited lifespan. Here, we created an immortalized melanocyte cell model—hTERT neonatal melanocytes—by retroviral transduction of human telomerase (hTERT) into primary cells. In addition to enhanced longevity (up to 35 doublings), physiologic marker expression (tyrosinase positive, fibroblast marker negative), and the ability to create melanosomes in 3D organotypic co-cultures, the cell line also showed expected levels of responses to several stimulators and inhibitors of melanogenesis. The inhibitors hydroquinone and kojic acid showed dose-dependent decreases in melanin content and the stimulators stem cell factor and latanoprost showed a muted response. In summary, immortalized melanocytes provided a versatile in vitro cell model for the study of skin toxicology and melanogenesis regulation.

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Presenter

Diana Douglas, headshot.

Diana Douglas, BS

Senior Biologist, ATCC

Diana Douglas is a Senior Biologist at ATCC. For the last four years, she has focused her research on the development of advanced biological models via CRISPR/Cas9 gene-editing technology. Previously, Ms. Douglas worked at the Baker Institute for Animal Health at Cornell University and the Dalton Cardiovascular Research Center at the University of Missouri, where her research focused on the mechanisms of necrotic cell death in heart disease. Ms. Douglas attended Truman State University where she obtained a Bachelor of Science in Biology.

hTERT kidney renal proximal tubular epithelial cells.

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