gms | German Medical Science

124. Kongress der Deutschen Gesellschaft für Chirurgie

Deutsche Gesellschaft für Chirurgie

01. - 04.05.2007, München

Restructuring skin equivalents with human outer root sheath cells and fibroblasts

Meeting Abstract

  • S. Liu - Department of Plastic Surgery and Hand Surgery, Burn Care Center, UK-SH Campus Lübeck, Lübeck, Germany
  • B. Bucsky - Department of Plastic Surgery and Hand Surgery, Burn Care Center, UK-SH Campus Lübeck, Lübeck, Germany
  • T. Egana - Department of Plastic Surgery and Hand Surgery, Burn Care Center, UK-SH Campus Lübeck, Lübeck, Germany
  • I. Wilcke - Department of Plastic Surgery and Hand Surgery, Burn Care Center, UK-SH Campus Lübeck, Lübeck, Germany
  • J. Lohmeyer - Department of Plastic Surgery and Hand Surgery, Burn Care Center, UK-SH Campus Lübeck, Lübeck, Germany
  • E. Bodo - UKSH / Campus Lübeck, Lübeck, Germany
  • S. Krüger - UKSH / Campus Lübeck, Lübeck, Germany
  • Z. Lu - UKSH / Campus Lübeck, Lübeck, Germany
  • corresponding author H.-G. Machens - Department of Plastic Surgery and Hand Surgery, Burn Care Center, UK-SH Campus Lübeck, Lübeck, Germany

Deutsche Gesellschaft für Chirurgie. 124. Kongress der Deutschen Gesellschaft für Chirurgie. München, 01.-04.05.2007. Düsseldorf: German Medical Science GMS Publishing House; 2007. Doc07dgch7608

Die elektronische Version dieses Artikels ist vollständig und ist verfügbar unter: http://www.egms.de/de/meetings/dgch2007/07dgch449.shtml

Veröffentlicht: 1. Oktober 2007

© 2007 Liu et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

Introduction: The bulge region, a portion of the outer root sheath (ORS), contains the hair follicle stem cells, which shows multipotency to differentiate into almost all epithelial cell types. To produce an improved quality skin equivalent (SE), we established a rapid, easy and effective three-dimensional SE model on the basis of human dermal fibroblasts, collagen-GAG matrix (DE) and human ORS cells (ORSCs).

Materials and methods: We dissected hair follicles from temporal scalp tissue after face-lift surgery and isolated ORSCs from the middle and lower parts of hair follicles. The ORSCs were primarily cultured and subcultured in KD-SFM for cell studies and seeded into collagen-GAG matrices. The matrices were seeded with human fibroblasts, cultured in DMEM with 10% FBS to form a dermal equivalent (DE). 24 h later, passage 1 ORSCs were seeded into the DE and cultured. 4 days later, the surface of the SEs was lifted to the air-liquid interface. One week after the air-liquid co-culture, Matrices were transplanted into bilateral 15 mm in diameter full-thickness wounds of athymic nu/nu mice. Wound tissue samples were harvested and examined by means of histology and immunohistochemistry.

Results: The isolated ORSCs had high proliferation ability. The expression of the immunofluorescence antibodies of CK15, CK19 and Integrin ß1 were all positive in the primary ORSCs. After passage 2, the expression of Integrin ß1 and CK15 decreased, while expression of CK19 was still positive. 2 weeks after transplantation, a fully developed, multi-layered and cornified epidermis layer could be observed in the matrices and stabilized after 4 weeks. The layers were stained positive both by antibody KL-1 and HLA-1, proving its human origin.

Discussion: This study gives methodological evidence that our isolation technique allows culturing of human ORSCs. The SE which we restructured with fibroblasts and OSRCs in vivo is able to form histological structures similar to human dermis and epidermis within 2 weeks.