title: Ex Vivo Prefabricated Rat Skin Flap Using Cell Sheets and an Arteriovenous Vascular Bundle.
abstract: BACKGROUND
  Recently, research on tissue-engineered skin substitutes have been active in plastic surgery, and significant development has been made in this area over the past several decades. However, a regenerative skin flap has not been developed that could provide immediate blood flow after transplantation. Here, we make a regenerative skin flap ex vivo that is potentially suitable for microsurgical transplantation in future clinical applications.
  
  
  METHODS
  In rats, for preparing a stable vascular carrier, a femoral vascular pedicle was sandwiched between collagen sponges and inserted into a porous chamber in the abdomen. The vascular bed was harvested 3 weeks later, and extracorporeal perfusion was performed. A green fluorescent protein positive epidermal cell sheet was placed on the vascular bed. After perfusion culture, the whole construct was harvested and fixed for morphological analyses.
  
  
  RESULTS
  After approximately 10 days perfusion, the epidermal cell sheet cornified sufficiently. The desquamated corneum was positive for filaggrin. The basement membrane protein laminin 332 and type 4 collagen were deposited on the interface area between the vascular bed and the epidermal cell sheet. Moreover, an electron microscopic image showed anchoring junctions and keratohyalin granules. These results show that we were able to produce native-like skin.
  
  
  CONCLUSIONS
  We have succeeded in creating regenerative skin flap ex vivo that is similar to native skin, and this technique could be applied to create various tissues in the future.
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Fujisawa
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Sekine
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sakurai
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Shimizu