Assembly of complete mouse embryo models from embryonic and induced stem cell types in vitro.
Nat Protoc
; 18(12): 3662-3689, 2023 Dec.
Article
em En
| MEDLINE
| ID: mdl-37821625
ABSTRACT
The interaction between embryonic and extraembryonic tissues is critical in natural mouse embryogenesis. Here, to enable such interaction in vitro, we describe a protocol to assemble a complete mouse embryo model using mouse embryonic stem cells and induced embryonic stem cells to express Cdx2 (or trophoblast stem cells) and Gata4 to reconstitute the epiblast, extraembryonic ectoderm and visceral endoderm lineages, respectively. The resulting complete embryo models recapitulate development from embryonic day 5.0 to 8.5, generating advanced embryonic and extraembryonic tissues that develop through gastrulation to initiate organogenesis to form a head and a beating heart structure as well as a yolk sac and chorion. Once the required stem cell lines are stably maintained in culture, the protocol requires 1 day to assemble complete embryo models and a further 8 days to culture them until headfold stages, although structures can be collected at earlier developmental stages as required. This protocol can be easily performed by researchers with experience in mouse stem cell culture, although they will benefit from knowledge of natural mouse embryos at early postimplantation stages.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Embrião de Mamíferos
/
Camadas Germinativas
Limite:
Animals
Idioma:
En
Revista:
Nat Protoc
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Reino Unido