Efficient propagation of single cells Accutase-dissociated human embryonic stem cells.
Mol Reprod Dev
; 75(5): 818-27, 2008 May.
Article
em En
| MEDLINE
| ID: mdl-18157870
ABSTRACT
Human embryonic stem cells (hESCs) hold great promise for cell-based therapies and drug screening applications. However, growing and processing large quantities of undifferentiated hESCs is a challenging task. Conventionally, hESCs are passaged as clusters, which can limit their growth efficiency and use in downstream applications. This study demonstrates that hESCs can be passaged as single cells using Accutase, a formulated mixture of digestive enzymes. In contrast to trypsin treatment, Accutase treatment does not significantly affect the viability and proliferation rate of hESC dissociation into single cells. Accutase-dissociated single cells can be separated by FACS and proliferate as fully pluripotent hESCs. An Oct4-eGFP reporter construct engineered into hESCs was used to monitor the pluripotency of hESCs passaged with Accutase. Compared to collagenase-passaged hESCs, Accutase-treated cultures contained a larger proportion of undifferentiated (Oct4-positive) cells. Additionally, Accutase-passaged undifferentiated hESCs could be grown as monolayers without the need for monitoring and/or selection for quality hESC colonies.
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Base de dados:
MEDLINE
Assunto principal:
Técnicas de Cultura de Células
/
Células-Tronco Pluripotentes
/
Proliferação de Células
/
Enzimas
/
Células-Tronco Embrionárias
Limite:
Humans
Idioma:
En
Revista:
Mol Reprod Dev
Assunto da revista:
BIOLOGIA MOLECULAR
/
MEDICINA REPRODUTIVA
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Estados Unidos