Effect of keratinocytes culture on the construction of fibrin-based human skin equivalents

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Published: Dec 13, 2022
DOI: https://doi.org/10.14456/sehs.2022.34
Keywords:
explant culture feeder layer fibrin gel human skin equivalents transepithelial electrical resistance

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Catalina Gaviria-Agudelo
Tissue Engineering and Cell Therapy Research Group, School of Medicine, University of Antioquia, Medellín, Colombia / European Research Institute for the Biology of Ageing, University Medical Center Groningen, Groningen, The Netherlands. Corresponding author's e-mail: [email protected]
Natalia Becerra
Tissue Engineering and Cell Therapy Research Group, School of Medicine, University of Antioquia, Medellín, Colombia
Luis Alfonso Correa
Tissue Engineering and Cell Therapy Research Group, School of Medicine, University of Antioquia, Medellín, Colombia / Dermatology Department, School of Medicine, University of Antioquia, Medellín, Colombia / Laboratorio Clínico VID, Congregación Mariana, Medellín, Colombia
Sergio Estrada
Tissue Engineering and Cell Therapy Research Group, School of Medicine, University of Antioquia, Medellín, Colombia
Luz Marina Restrepo
Tissue Engineering and Cell Therapy Research Group, School of Medicine, University of Antioquia, Medellín, Colombia / Medical Research Institute, School of Medicine, University of Antioquia, Medellín, Colombia

Abstract

Human skin equivalents are in vitro model constructed with human skin cells. The function and morphology of primary cells are particularly dependent on the cell isolation and culturing method, crucial in achieving reliable skin tissues. In this study, we show preliminary findings on the effect of the source of keratinocytes on the generation of 3D skin models. Two approaches were used to obtain keratinocytes: explant culture and a feeder layer of mouse fibroblasts. Skin samples were taken from three patients and processed by both methods. For the construction of HSEs, explant culture and feeder layer-derived keratinocytes were seeded on top of a fibroblasts-populated fibrin matrix. The histology and expression of epidermal markers were assessed by hematoxylin-eosin staining and immunohistochemistry, respectively. The integrity of the epidermal barrier was examined by measuring the transepithelial electrical resistance. To a greater or lesser extent, both methods produced HSEs where keratinocytes were able to stratify and express epidermal differentiation markers. The integrity of tight junctions (protein complexes that form the epidermal barrier) was enhanced in models composed of passage 1 cells. Feeder layer-derived keratinocytes generated HSEs with a healthier, thicker, and more properly stratified epidermis, improving the histological features, when compared to explant culture-derived models.

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How to Cite
Gaviria-Agudelo, C., Becerra, N., Alfonso Correa, L., Estrada, S., & Marina Restrepo, L. (2022). Effect of keratinocytes culture on the construction of fibrin-based human skin equivalents. Science, Engineering and Health Studies, 16, 22030008. https://doi.org/10.14456/sehs.2022.34
Issue
Volume 16, 2022
Section
Biological sciences
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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

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