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. 2022 Oct 22;391(1):173–188. doi: 10.1007/s00441-022-03698-7

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Serum-free 3D-engineered skin structure. a–c Collagen IV (ColIV, a basement membrane marker) and pan-cytokeratin (panCK) immunofluorescence staining showed higher ColIV deposition in the basement membrane when fibroblasts were seeded on top (CT = cells on top) of the hydrogel. The amount of cytokeratin in the epidermis was similar whether the fibroblasts were seeded on top or within the hydrogel (CW = cells within). d–f CK5 (a marker for interfollicular stem and progenitor keratinocytes) and CK10 (a differentiation marker) immunofluorescence staining confirmed CK5 expression in basal keratinocytes, when fibroblasts were seeded on top or within the hydrogel, although at a lower level, when compared to native skin. Basal keratinocytes did not express CK10, similar to native skin, regardless of whether fibroblasts were seeded on top or within the hydrogel. EPI, epidermis; DERM, dermis; and white dotted line, dermal/epidermal junction (scale bar 100 µm). e Integrated density quantitation for ColIV and panCK measured using FIJI software (n = 3). f Integrated density quantitation for CK5 and CK10 (* = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001) (n = 4)