title: Establishment of Immortalized Primary Human Foreskin Keratinocytes and Their Application to Toxicity Assessment and Three Dimensional Skin Culture Construction.
abstract: In spite of frequent usage of primary human foreskin keratinocytes (HFKs) in the study of skin biology, senescence-induced blockage ofin vitroproliferation 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 ofin vitroskin disease models but also for prediction of potential toxicities of various cosmetic chemicals.
authors.affiliation: College of Pharmacy, Dongguk University, Goyang 10326, Republic of Korea.
authors.forename: Moonju
authors.initials: M
authors.lastname: Choi
authors.affiliation: College of Pharmacy, Dongguk University, Goyang 10326, Republic of Korea.
authors.forename: Minkyung
authors.initials: M
authors.lastname: Park
authors.affiliation: R&D Institute, Biosolution Co., Ltd., Seoul 01811, Republic of Korea.
authors.forename: Suhyon
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Urology, Dongguk University Ilsan Hospital, Dongguk University College of Medicine, Goyang 10326, Republic of Korea.
authors.forename: Jeong Woo
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Department of Urology, Seoul Metropolitan Government-Seoul National University (SMG-SNU) Boramae Medical Center, Seoul 07061, Republic of Korea.
authors.forename: Min Chul
authors.initials: MC
authors.lastname: Cho
authors.affiliation: College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Minsoo
authors.initials: M
authors.lastname: Noh
authors.affiliation: College of Pharmacy, Dongguk University, Goyang 10326, Republic of Korea.
authors.forename: Choongho
authors.initials: C
authors.lastname: Lee
keywords: Human foreskin keratinocyte; Immortalization; Three-dimensional skin culture; Toxicity assessment