Efficient Derivation of Pluripotent Stem Cells from SiRNA-mediated Cdx2-deficient Mouse Embryos

Overview

Journal Stem Cells Dev

Date 2010 Jun 12

PMID 20536317

Citations 5

Authors

Guangming Wu

Luca Gentile

Jeong Tae Do

Tobias Cantz

Julien Sutter

Katherina Psathaki

Marcos J Arauzo-Bravo

Claudia Ortmeier

Hans R Scholer

Affiliations

Soon will be listed here.

Abstract

In the early mammalian embryo, lineage separation of and subsequent crosstalk between the trophectoderm (TE) and inner cell mass (ICM) are required to support further development. Previous studies have shown that the homeobox transcription factor Cdx2 is required for TE differentiation and that lack of Cdx2 expression causes death of embryos at the peri-implantation stage. In this study, we effectively eliminated Cdx2 transcripts by microinjection of siRNA into embryos and evaluated the effect on efficiency of deriving embryonic stem cells (ESCs). By this approach, we successfully created nonviable embryos similar to reported knockout embryos. Accordingly, the efficiency of ESC derivation dropped from 19.1% in control blastocysts to 2% in Cdx2-deficient blastocysts, indicating loss of pluripotency in the ICM. Strikingly, when 8-cell stage embryos were cultured under ESC culture conditions before lineage separation, fully functional pluripotent stem cell lines were obtained, with efficiency even greater than that for control embryos. These results demonstrate that Cdx2 plays an essential role within the microenvironment created by the TE to support ICM pluripotency but that the ESC culture system, with mouse embryonic fibroblasts, could rescue the pluripotent cell population for efficient ESC derivation.

Citing Articles

Leptin treatment of in vitro cultured embryos increases outgrowth rate of inner cell mass during embryonic stem cell derivation.

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Establishment of bovine embryonic stem cells after knockdown of CDX2.

Wu X, Song M, Yang X, Liu X, Liu K, Jiao C Sci Rep. 2016; 6:28343.

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Establishment of totipotency does not depend on Oct4A.

Wu G, Han D, Gong Y, Sebastiano V, Gentile L, Singhal N Nat Cell Biol. 2013; 15(9):1089-97.

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A potential use of embryonic stem cell medium for the in vitro culture of preimplantation embryos.

Gelber K, Tamura A, Alarcon V, Marikawa Y J Assist Reprod Genet. 2011; 28(8):659-68.

PMID: 21617931 PMC: 3170111. DOI: 10.1007/s10815-011-9587-8.

Initiation of trophectoderm lineage specification in mouse embryos is independent of Cdx2.

Wu G, Gentile L, Fuchikami T, Sutter J, Psathaki K, Esteves T Development. 2010; 137(24):4159-69.

PMID: 21098565 PMC: 2990207. DOI: 10.1242/dev.056630.

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