Effects of culture media on gene expression in reconstructed human epidermis and THP-1 monocytes for skin sensitization evaluation in co-culture systems

被引：0

作者：

Sugimoto-Sawada, Y. ^{[1
]}

Yamashiro, M. ^{[1
]}

Kono, M. ^{[2
]}

Ikeda, H. ^{[2
]}

Itagaki, H. ^{[3
]}

Iijima, K. ^{[4
,5
]}

机构：

[1] Yokohama Natl Univ, Grad Sch Engn Sci, 79-5 Tokiwadai,Hodogaya Ku, Yokohama 2408501, Japan

[2] Mandom Corp, 5-12 Junikencho,Chuo Ku, Osaka, Osaka 5408530, Japan

[3] ITACS Consulting, 5-11-19-2504 Minamidai,Minami Ku, Sagamihara 2520314, Japan

[4] Yokohama Natl Univ, Fac Engn, 79-5 Tokiwadai,Hodogaya Ku, Yokohama 2408501, Japan

[5] Yokohama Natl Univ, Inst Adv Sci, 79-5 Tokiwadai, Hodogaya ku, Yokohama 2408501, Japan

来源：

TOXICOLOGY IN VITRO | 2025年 / 106卷

关键词：

Skin sensitization; Alternative method; Reconstructed human epidermis (RhE) model; Gene expression; Co-culture system; Crosstalk; FACTOR BINDING PROTEIN-3; DENDRITIC CELLS; MICROPHYSIOLOGICAL SYSTEMS; S100; PROTEINS; KERATINOCYTES; ACTIVATION; TRANSCRIPTION; RECEPTOR; LINE; GRANULYSIN;

D O I：

10.1016/j.tiv.2025.106035

中图分类号：

R99 [毒物学（毒理学）];

学科分类号：

100405 ;

摘要：

Co-culture with reconstituted epidermis formed by normal human epidermal keratinocytes (RhE) increases the expression of the skin sensitization markers CD54 and CD86 on the human monocytic leukemia cell line THP-1 without chemicals. Therefore, we investigated the effects of culture media [RPMI1640 for RhE; keratinization induction (KI) medium for THP-1], co-culture, and the responses to the skin sensitizer 2,4-dinitrochlorobenzene (DNCB) on gene expression in mono- and cocultures of RhE and THP-1 cells. Microarray and pathway analyses revealed that in mono-RhE, RPMI medium induced epidermal differentiation-related genes, whereas in monoculture THP-1 cells, KI medium upregulated inflammation-related genes. Surprisingly, the medium composition had a more significant impact than co-culture in both cells. However, crosstalk between RhE and THP-1 cells was observed upon DNCB exposure by comparing the differentially expressed gene sets. DNCB-treated THP-1 cells showed increased expression of NR4A1, NR4A2, NR4A3, SIK1, and HMOX1 in co-culture than in monoculture, and these gene expression patterns were confirmed by real-time RT-PCR. It has been suggested that danger signals from RhE, in response to DNCB, enhance the expression of these genes in THP-1 cells. We clarified the effects of the medium and co-culture and proposed these five genes as potential markers for skin sensitization evaluation.

引用

页数：23

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