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Transduction of β3 integrin subunit cDNA confers on human keratinocytes the ability to adhere to gelatin

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Abstract

αvβ3 is a multiligand integrin receptor that interacts with fibrinogen (FG), fibrin (FB), fibronectin (FN), vitronectin (VN), and denatured collagen. We previously reported that cultured normal human keratinocytes, like in vivo keratinocytes, do not express αvβ3 on the cell surface, and do not adhere to and migrate on FG and FB. Furthermore, we reported that human keratinocytes transduced with β3 integrin subunit cDNA by a retrovirus-mediated transduction method express αvβ3 on the cell surface and adhere to FG, FB, FN, and VN significantly compared with β-galactosidase (β-gal) cDNA-transduced keratinocytes (control). In this study, we determined whether these β3 integrin subunit cDNA-transduced keratinocytes or normal human keratinocytes adhere to denatured collagen (gelatin) using a 1 h cell adhesion assay. β3 cDNA-transduced keratinocytes adhered to gelatin, whereas no significant adhesion was observed with the control cells (β-gal cDNA-transduced keratinocytes and normal human keratinocytes). The adhesion to gelatin was inhibited by LM609, a monoclonal antibody to αvβ3, and RGD peptides but not by normal mouse IgG1 nor RGE peptides. Thus, transduction of β3 integrin subunit cDNA confers on human keratinocytes the ability to adhere to denatured collagen (gelatin) as well as to FG, FB, VN, and FN. Otherwise, normal human keratinocytes do not adhere to gelatin. These data support the idea that β3 cDNA-transduced human keratinocytes can be a good material for cultured epithelium to achieve better take rate with acute or chronic wounds, in which FG, FB, and denatured collagen are abundantly present.

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Abbreviations

KBM:

Keratinocyte basal medium

KGM:

Keratinocyte growth medium

PS:

Penicillin and streptomycin

β-gal:

β-Galactosidase

GEL:

Gelatin

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Acknowledgments

Funding of this work was provided by Research Project Grant (13-204) from Kawasaki Medical School, SHISEIDO Grant for Skin Aging Research (SRG#10048), JSPS.KAKENHI (15390543) to M. Kubo; and by National Institute of Health Grant (AG10143) to R. A. F. Clark.

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Authors and Affiliations

  1. Department of Plastic and Reconstructive Surgery, Kawasaki Medical School, 577 Matsushima, Kurashiki City, Okayama, 701-0192, Japan

    Miyoko Kubo, Ying Zhao & Takahiko Moriguchi

  2. Director of Center for Tissue Engineering, Department of Biomedical Engineering, Dermatology and Medicine, School of Medicine, SUNY at Stony Brook, HSC T-16,060, Stony Brook, NY, 11794-8165, USA

    Richard A. F. Clark

  3. Department of Oral Biology and Pathology, School of Dental Medicine, SUNY at Stony Brook, Stony Brook, NY, 11794-8702, USA

    Anne B. Katz & Lorne B. Taichman

  4. Deparment of Pathology, Mudanjiang Medical University, No.3 Tongxiang Street, Aimin District, Mudanjiang City, Heilongjiang Province, 157011, China

    Zaishun Jin

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  1. Miyoko Kubo
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Correspondence to Miyoko Kubo.

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Kubo, M., Clark, R.A.F., Katz, A.B. et al. Transduction of β3 integrin subunit cDNA confers on human keratinocytes the ability to adhere to gelatin. Arch Dermatol Res 299, 13–24 (2007). https://doi.org/10.1007/s00403-006-0718-5

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