Artigo de peri??dico
Phenotype changes of oral epithelial stem cells after in vitro culture
Registro en:
1806-8324
34
10.1590/1807-3107bor-2020.vol34.0033
41.85
72.00
Autor
DALTOE, FELIPE P.
OLIVEIRA, N??LIO A.J. de
PERONI, CIBELE N.
SHARPE, PAUL T.
MANTESSO, ANDREA
Resumen
The aim of our study was to isolate populations of
keratinocyte stem cells based on the expression of cell surface markers
and to investigate whether the culture could affect their phenotype.
keratinocytes from human oral mucosa were sorted based on the
expression of the epithelial stem cell markers p75NTR and CD71. We
also examined the co-expression of other epithelial stem markers such
as integrins ??1 and ??6 and their stem cell-like proprieties in in vitro
assays. Three passages after being sorted by MACS, more than 93%
of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of
the p75NTR-ve gained it. Within the small population of the p75NTR+ve
cells, 88% co-expressed other epithelial stem cell markers such as
integrins ??1 and ??6, while only 28% of p75NTR-ve cells co-expressed
these markers. These results were confirmed by sorting cells by
FACS. Additionally, when double staining was used for sorting cells,
99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells
expressed both integrins, but just one week after culture, only 1.74%
of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32%
still expressed CD71. Similar results were obtained when co-culturing
p75NTR+ve and p75NTR-ve populations before analysis. Our results
suggest that phenotype changes may be part of an intrinsic cellular
mechanism to conserve levels of protein expression as they may
found in the human body. In addition, in vitro culture may not offer
ideal conditions for epithelial stem cell maintenance due to phenotype
changes under standard culture conditions. Funda????o de Amparo ?? Pesquisa do Estado de S??o Paulo (FAPESP) Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior (CAPES) FAPESP: 08/11641-9 CAPES: 2586-11-8