Establishment of Immortalized Primary Human Foreskin Keratinocytes and Their Application to Toxicity Assessment and Three Dimensional Skin Culture Construction

• Published in: Biomolecules & therapeutics Vol. 25; no. 3; pp. 296 - 307
• Main Authors: Choi, Moonju, Park, Minkyung, Lee, Suhyon, Lee, Jeong Woo, Cho, Min Chul, Noh, Minsoo, Lee, Choongho
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Society of Applied Pharmacology 01.05.2017; 한국응용약물학회
• Subjects: Original; 약학; Human foreskin keratinocyte; Three-dimensional skin culture; Toxicity assessment; Immortalization
• ISSN: 1976-9148; 2005-4483; 1976-9148; 2005-4483
• DOI: 10.4062/biomolther.2017.043

In spite of frequent usage of primary human foreskin keratinocytes (HFKs) in the study of skin biology, senescence-induced blockage of proliferation has been a big hurdle for their effective utilization. In order to overcome this passage limitation, we first isolated ten HFK lines from circumcision patients and successfully immortalized four of them via a retroviral transduction of high-risk human papillomavirus (HPV) E6 and E7 oncogenes. We confirmed expression of a keratinocyte marker protein, keratin 14 and two viral oncoproteins in these immortalized HFKs. We also observed their robust responsiveness to various exogenous stimuli, which was evidenced by increased mRNA expression of epithelial differentiation markers and pro-inflammatory genes in response to three reactive chemicals. In addition, their applicability to cytotoxicity assessment turned out to be comparable to that of HaCaT cells. Finally, we confirmed their differentiation capacity by construction of well-stratified three dimensional skin cultures. These newly established immortalized HFKs will be valuable tools not only for generation of skin disease models but also for prediction of potential toxicities of various cosmetic chemicals.