Back to Search
Start Over
Tracking keratinocytes and melanocytes using carboxyfluorescein hydroxysuccinimidyl ester staining
- Source :
- PLoS ONE, Vol 14, Iss 8, p e0221878 (2019), PLoS ONE
- Publication Year :
- 2019
- Publisher :
- Public Library of Science (PLoS), 2019.
-
Abstract
- 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. Funding Agencies|Region Ostergotland [LIO-695901]
- Subjects :
- Keratinocytes
Physiology
Imaging Techniques
Cell Survival
Cell- och molekylärbiologi
Science
Fluorescent Antibody Technique
Succinimides
Surgical and Invasive Medical Procedures
Research and Analysis Methods
Epithelium
Fluorescence Microscopy
Animal Cells
Tissue Repair
Fluorescence Imaging
Medicine and Health Sciences
Humans
Chromatophores
Fluorescent Dyes
Staining
Microscopy
Wound Healing
Chemical Compounds
Biology and Life Sciences
Cell Staining
Light Microscopy
Epithelial Cells
Esters
Cell Biology
Fluoresceins
Immunohistochemistry
Skin Grafting
Chemistry
Biological Tissue
Microscopy, Fluorescence
Specimen Preparation and Treatment
Physical Sciences
Melanocytes
Medicine
Cellular Types
Anatomy
Physiological Processes
Plastic Surgery and Reconstructive Techniques
Cell and Molecular Biology
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 14
- Issue :
- 8
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.pmid.dedup....fd56f82ff61b7c7343708c3bcd54020e