ROCK inhibitor combined with Ca2+ controls the myosin II activation and optimizes human nasal epithelial cell sheets.
Sci Rep
; 10(1): 16853, 2020 10 08.
Article
em En
| MEDLINE
| ID: mdl-33033339
ABSTRACT
The proliferation and differentiation of cultured epithelial cells may be modified by Rho-associated kinase (ROCK) inhibition and extracellular Ca2+ concentration. However, it was not known whether a combination would influence the behavior of cultured epithelial cells through changes in the phosphorylation of non-muscle myosin light chain II (MLC). Here we show that the combination of ROCK inhibition with Ca2+ elevation regulated the phosphorylation of MLC and improved both cell expansion and cell-cell adhesion during the culture of human nasal mucosal epithelial cell sheets. During explant culture, Ca2+ enhanced the adhesion of nasal mucosal tissue, while ROCK inhibition downregulated MLC phosphorylation and promoted cell proliferation. During cell sheet culture, an elevation of extracellular Ca2+ promoted MLC phosphorylation and formation of cell-cell junctions, allowing the harvesting of cell sheets without collapse. Moreover, an in vitro grafting assay revealed that ROCK inhibition increased the expansion of cell sheets three-fold (an effect maintained when Ca2+ was also elevated), implying better wound healing potential. We suggest that combining ROCK inhibition with elevation of Ca2+ could facilitate the fabrication of many types of cell graft.
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Base de dados:
MEDLINE
Assunto principal:
Piridinas
/
Adesão Celular
/
Diferenciação Celular
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Cálcio
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Técnicas de Cultura de Células
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Miosina Tipo II
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Proliferação de Células
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Inibidores Enzimáticos
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Células Epiteliais
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Quinases Associadas a rho
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article