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Abdallah, Sallam
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Lagerwall, C., Shahin, H., Abdallah, S., Steinvall, I., Elmasry, M., Sjöberg, F. & El-Serafi, A. T. (2021). Xeno-free workflow exhibits comparable efficiency and quality of keratinocytes isolated from human skin biopsies. Regenerative Therapy, 18, 401-407
Open this publication in new window or tab >>Xeno-free workflow exhibits comparable efficiency and quality of keratinocytes isolated from human skin biopsies
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2021 (English)In: Regenerative Therapy, ISSN 2352-3204, Vol. 18, p. 401-407Article in journal (Refereed) Published
Abstract [en]

Introduction Regenerative solutions of the skin represent a hope for burn victims with extensive skin loss and chronic wound patients. The development of xeno-free workflow is crucial for clinical application in compliance with the directives of the European Medicines Agency. This study aimed at evaluating the outcome of the xeno-free isolation workflow of keratinocytes from human skin biopsy. Methods Skin biopsies were obtained from volunteers. The epidermis was digested with TrypLE™ Select, which was deactivated by dilution or with trypsin, deactivated by media with fetal bovine serum. Freshly isolated cells were compared for total cell number, viability, activity of caspase 3, gene expression and the presence of the keratinocyte surface markers cytokeratin 14. The cells were cultured in xeno-free conditions for one week and characterized regarding the number and viability as well as the metalloproteinase secretion. Results The number of obtained cells was similar in both workflows. The cell viability was less in the TrypLE group, with slight reduction of the cell surface marker cytokeratin 14. Caspase 3 activity was comparable as well as the gene expression of the apoptotic markers BAX, BCL2 and SLUG, as well as the keratinocyte markers cytokeratin 14, stratifin and filaggrin. Upon culture, the number of keratinocytes, their viability and secretion of matrix metalloproteinases 1 and 10 were equal in both groups. Conclusion This study reports the possibility of isolating functioning and viable keratinocytes through a xeno-free workflow for clinical application.

Place, publisher, year, edition, pages
Elsevier, 2021
Keywords
Keratinocytes, Regenerative medicine, European medicines agency, Xeno-free, TrypLE, Trypsin
National Category
Surgery
Identifiers
urn:nbn:se:liu:diva-179790 (URN)10.1016/j.reth.2021.09.005 (DOI)000703088900050 ()
Note

Funding agencies: Centre for Advanced Medical Product, Sweden; Hand and Plastic Surgery Department, Linköping University Hospital, Region Östergötland, Sweden.

Available from: 2021-10-01 Created: 2021-10-01 Last updated: 2023-11-09Bibliographically approved
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