Generation of nuclear transfer-derived pluripotent ES cells from cloned Cdx2-deficient blastocysts.
The derivation of embryonic stem (ES) cells by nuclear transfer holds great promise for research and therapy but involves the destruction of cloned human blastocysts. Proof of principle experiments have shown that 'customized' ES cells derived by nuclear transfer (NT-ESCs) can be used to correct immunodeficiency in mice. Importantly, the feasibility of the approach has been demonstrated recently in humans, bringing the clinical application of NT-ESCs within reach. Altered nuclear transfer (ANT) has been proposed as a variation of nuclear transfer because it would create abnormal nuclear transfer blastocysts that are inherently unable to implant into the uterus but would be capable of generating customized ES cells. To assess the experimental validity of this concept we have used nuclear transfer to derive mouse blastocysts from donor fibroblasts that carried a short hairpin RNA construct targeting Cdx2. Cloned blastocysts were morphologically abnormal, lacked functional trophoblast and failed to implant into the uterus. However, they efficiently generated pluripotent embryonic stem cells when explanted into culture.
Meissner
Alexander
A
Whitehead Institute and Department of Biology, Massachusetts Institute of Technology, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA.
Jaenisch
Rudolf
R
D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism*; D000071616:CDX2 Transcription Factor; D018929:Cell Culture Techniques; D002470:Cell Survival; D010064:Embryo Implantation; D004624:Embryo Transfer; D005260:Female; D018398:Homeodomain Proteins / Q000235:genetics; D051379:Mice; D053652:Nuclear Transfer Techniques; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011555:Pseudopregnancy; D015203:Reproducibility of Results; D035845:Research Embryo Creation; D014157:Transcription Factors / Q000172:deficiency* / Q000235:genetics
D000071616:CDX2 Transcription Factor; C487514:Cdx2 protein, mouse; D018398:Homeodomain Proteins; D014157:Transcription Factors