IGF-1R deficiency in human keratinocytes disrupts epidermal homeostasis and stem cell maintenance.


Bioscience & Technology Development Center, FUJIFILM Corporation, 577 Kaisei, Kanagawa 258-8577, Japan; Department of Stem Cell Biology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan. Electronic address: [Email]


BACKGROUND : Epidermal stem cells (ESCs) are keratinocytes that reside in the basal layer of the epidermis and mediate epidermal homeostasis. Insulin-like growth factor 1 (IGF-1) signaling through its receptor (IGF-1R) has been identified as an important regulator in rodent skin development and differentiation. However, the role of IGF-1/IGF-1R signaling in human keratinocytes is not yet well understood.
OBJECTIVE : This study aimed to clarify the role of IGF-1/IGF-1R signaling in human epidermal homeostasis.
METHODS : IGF-1R specific knockout (KO) HaCaT keratinocytes were generated by CRISPR-Caspase-9-mediated non-homologous end joining frame-shift mutations. Further, the behavior of these keratinocytes in epidermal homeostasis was investigated using reconstructed epidermis and human skin equivalents.
RESULTS : IGF-1R KO HaCaT keratinocytes were successfully established and produced thin epidermis in three-dimensional culture models. Keratin10-positive cells were frequently found in the basal layer of the reconstructed epidermis.
CONCLUSIONS : IGF-1/IGF-1R signaling was demonstrated to play a key role in maintaining human epidermal homeostasis. This method provides a new framework to investigate gene function in human epidermal homeostasis.


CRISPR-Cas9,Human skin equivalent,IGF-1/IGF-1R signaling,Keratinocyte,

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