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Tracking keratinocytes and melanocytes using carboxyfluorescein hydroxysuccinimidyl ester staining
Linköping University, Department of Clinical and Experimental Medicine, Division of Surgery, Orthopedics and Oncology. Linköping University, Faculty of Medicine and Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine, Division of Surgery, Orthopedics and Oncology. Linköping University, Faculty of Medicine and Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Medicine and Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine, Division of Surgery, Orthopedics and Oncology. Linköping University, Faculty of Medicine and Health Sciences.
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2019 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 14, no 8, article id e0221878Article in journal (Refereed) Published
Abstract [en]

Introduction The treatment of burn wounds and hypopigmentation conditions often require autologous transplantation of keratinocytes and melanocytes. Tracking transplanted cells to ascertain their contribution to tissue recapitulation presents a challenge. This study demonstrates a methodology based on passive staining with carboxyfluorescein hydroxysuccinimidyl ester ( CFSE) that enables localization of cells in tissue sections to investigate the fate of transplanted cells in wound re-epithelialisation. Methods Viability and migration of CFSE-stained keratinocytes and melanocytes were investigated using viability staining and scratch assays, while proliferation of cells was measured using flow cytometry. In addition, CFSE-stained keratinocytes and melanocytes were transplanted to a human ex vivo wound model, either in suspension, or with the aid of macroporous gelatine microcarriers. Wounds were analysed seven, 14 and 21 days post transplantation using cryosectioning and fluorescence microscopy. Sections from wounds with transplanted co-cultured keratinocytes and melanocytes were stained for pancytokeratin to distinguish keratinocytes. Results CFSE-staining of keratinocytes and melanocytes did not affect the viability, migration or proliferation of the cells. Transplanted cells were tracked in ex vivo wounds for 21 days, illustrating that the staining had no effect on wound re-epithelialisation. In conclusion, this study presents a novel application of CFSE-staining for tacking transplanted primary human keratinocytes and melanocytes.

Place, publisher, year, edition, pages
PUBLIC LIBRARY SCIENCE , 2019. Vol. 14, no 8, article id e0221878
National Category
Cell and Molecular Biology
Research subject
Disaster Medicine
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URN: urn:nbn:se:liu:diva-160616DOI: 10.1371/journal.pone.0221878ISI: 000485058200063PubMedID: 31465496OAI: oai:DiVA.org:liu-160616DiVA, id: diva2:1362628
Note

Funding Agencies|Region Ostergotland [LIO-695901]

Available from: 2019-10-21 Created: 2019-10-21 Last updated: 2019-12-05

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Lönnqvist, SusannaJunker, JohanSedell, MariaNyman, ErikaKratz, Gunnar
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Division of Surgery, Orthopedics and OncologyFaculty of Medicine and Health SciencesDepartment of Clinical and Experimental MedicineDepartment of Hand and Plastic Surgery
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