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 cocultured
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.