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. 2014 Mar 17;22(2):246–255. doi: 10.1111/wrr.12154

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© 2014 The Authors. Wound Repair and Regeneration published by Wiley Periodicals, Inc. on behalf of Wound Healing Society.

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Histological and immunohistochemical staining shows differences between BLCC, Fibroblast matrix, and Epithelium only constructs. Hematoxylin and eosin (H&E) (A–C); Cytokeratin 1/10 (D–F); Collagen type IV (G–I); and Ki67 (J–L) show functional and morphological differences between constructs. Differentiated keratinocytes were only identified in the BLCC (D–F). Collagen type IV, indicating basement membrane deposition, and Ki67 immunostaining for proliferating cells were found at the interface of the dermal and epidermal layers in the BLCC, but not in Fibroblast matrix or Epithelium only constructs (G–I, J–L). Dotted lines in (A), (D), (G), and (J) demarcate the dermal–epidermal interface in the BLCC. Day 20 results shown. Each image is representative of n = 3 samples per group. BLCC, bilayered living cellular construct.