Induction of the G2/M transition stabilizes haploid embryonic stem cells.
Development
; 141(20): 3842-7, 2014 Oct.
Article
em En
| MEDLINE
| ID: mdl-25252944
ABSTRACT
The recent successful establishment of mouse parthenogenetic haploid embryonic stem cells (phESCs) and androgenetic haploid ESCs (ahESCs) has stimulated genetic research not only in vitro but also in vivo because of the germline competence of these cell lines. However, it is difficult to maintain the haploid status over time without a frequent sorting of the G1 phase haploid ESCs by fluorescence-activated cell sorting (FACS) at short intervals, because haploid cells tend to readily self-diploidize. To overcome this spontaneous diploid conversion, we developed a phESC culture condition using a small molecular inhibitor of Wee1 kinase to regulate the cell cycle by accelerating the G2/M phase transition and preventing re-entry into extra G1/S phase. Here, we demonstrate that, under this condition, phESCs maintained the haploid status for at least 4â
weeks without the need for FACS. This method will greatly enhance the availability of these cells for genetic screening.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Regulação da Expressão Gênica no Desenvolvimento
/
Células-Tronco Embrionárias
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Pontos de Checagem da Fase G2 do Ciclo Celular
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Haploidia
Limite:
Animals
Idioma:
En
Revista:
Development
Ano de publicação:
2014
Tipo de documento:
Article