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Reprogramming of two somatic nuclei in the same ooplasm leads to pluripotent embryonic stem cells

Reprogramming of two somatic nuclei in the same ooplasm leads to pluripotent embryonic stem cells
Reprogramming of two somatic nuclei in the same ooplasm leads to pluripotent embryonic stem cells
The conversion of the nuclear program of a somatic cell from a differentiated to an undifferentiated state can be accomplished by transplanting its nucleus to an enucleated oocyte (SCNT) in a process termed ‘reprogramming'. This process achieves pluripotency and occasionally also totipotency. Exploiting the obstacle of tetraploidy to full development in mammals, we show that mouse ooplasts transplanted with two somatic nuclei simultaneously (double SCNT) support preimplantation development and derivation of novel tetraploid SCNT embryonic stem cells (tNT-ESCs). Although the double SCNT embryos do not recapitulate the expression pattern of the pluripotency-associated gene Oct4 in fertilized embryos, derivative tNT-ESCs have characteristics of genuine pluripotency: in vitro they differentiate into neurons, cardiomyocytes and endodermal cells; in vivo, tNT-ESCs form teratomas, albeit at reduced rates compared to diploid counterparts. Global transcriptome analysis revealed only few specific alterations, e.g. in the quantitative expression of gastrulation-associated genes. In conclusion, we have shown that the oocyte's reprogramming capacity is in excess of a single nucleus and that double nucleus transplanted embryos and derivative ESCs are very similar to their diploid counterparts. These results have key implications for reprogramming studies based on pluripotency: while reprogramming in the tetraploid state was known from fusion-mediated reprogramming and from fetal and adult hepatocyte-derived iPS cells, we have now accomplished it with enucleated oocytes
nuclear transfer techniques, nuclear reprogramming, alleles, embryonic stem cells, tetraploidy, oct4, pou5f1
1066-5099
2343-2353
Pfeiffer, Martin J.
00e1c90a-e114-4f7d-b7e6-f0d49f447cb2
Esteves, Telma C.
038b596d-42fd-4c47-9c69-42bc227eeda9
Balbach, Sebastian T.
e2461a46-58ac-4c57-bc9f-f0ef59b18d03
Araúzo-Bravo, Marcos J.
6b27faaa-4924-4a4a-b1f4-9dde25d12530
Stehling, Martin
cd1db61d-1ca8-48f3-9cb1-eec1c24a3a28
Jauch, Anna
4878ab3f-1adc-4eb8-b038-d1f8d51c3e7a
Houghton, Franchesca D.
53946041-127e-45a8-9edb-bf4b3c23005f
Schwarzer, Caroline
d8e0cd47-6aae-4efc-8f90-e74731d84af9
Boiani, Michele
1979e7de-72f1-44fc-96c4-e1729afcbc6f
Pfeiffer, Martin J.
00e1c90a-e114-4f7d-b7e6-f0d49f447cb2
Esteves, Telma C.
038b596d-42fd-4c47-9c69-42bc227eeda9
Balbach, Sebastian T.
e2461a46-58ac-4c57-bc9f-f0ef59b18d03
Araúzo-Bravo, Marcos J.
6b27faaa-4924-4a4a-b1f4-9dde25d12530
Stehling, Martin
cd1db61d-1ca8-48f3-9cb1-eec1c24a3a28
Jauch, Anna
4878ab3f-1adc-4eb8-b038-d1f8d51c3e7a
Houghton, Franchesca D.
53946041-127e-45a8-9edb-bf4b3c23005f
Schwarzer, Caroline
d8e0cd47-6aae-4efc-8f90-e74731d84af9
Boiani, Michele
1979e7de-72f1-44fc-96c4-e1729afcbc6f

Pfeiffer, Martin J., Esteves, Telma C., Balbach, Sebastian T., Araúzo-Bravo, Marcos J., Stehling, Martin, Jauch, Anna, Houghton, Franchesca D., Schwarzer, Caroline and Boiani, Michele (2013) Reprogramming of two somatic nuclei in the same ooplasm leads to pluripotent embryonic stem cells. Stem Cells, 31 (11), 2343-2353. (doi:10.1002/stem.1497). (PMID:23922292)

Record type: Article

Abstract

The conversion of the nuclear program of a somatic cell from a differentiated to an undifferentiated state can be accomplished by transplanting its nucleus to an enucleated oocyte (SCNT) in a process termed ‘reprogramming'. This process achieves pluripotency and occasionally also totipotency. Exploiting the obstacle of tetraploidy to full development in mammals, we show that mouse ooplasts transplanted with two somatic nuclei simultaneously (double SCNT) support preimplantation development and derivation of novel tetraploid SCNT embryonic stem cells (tNT-ESCs). Although the double SCNT embryos do not recapitulate the expression pattern of the pluripotency-associated gene Oct4 in fertilized embryos, derivative tNT-ESCs have characteristics of genuine pluripotency: in vitro they differentiate into neurons, cardiomyocytes and endodermal cells; in vivo, tNT-ESCs form teratomas, albeit at reduced rates compared to diploid counterparts. Global transcriptome analysis revealed only few specific alterations, e.g. in the quantitative expression of gastrulation-associated genes. In conclusion, we have shown that the oocyte's reprogramming capacity is in excess of a single nucleus and that double nucleus transplanted embryos and derivative ESCs are very similar to their diploid counterparts. These results have key implications for reprogramming studies based on pluripotency: while reprogramming in the tetraploid state was known from fusion-mediated reprogramming and from fetal and adult hepatocyte-derived iPS cells, we have now accomplished it with enucleated oocytes

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More information

e-pub ahead of print date: 7 August 2013
Published date: November 2013
Keywords: nuclear transfer techniques, nuclear reprogramming, alleles, embryonic stem cells, tetraploidy, oct4, pou5f1
Organisations: Human Development & Health

Identifiers

Local EPrints ID: 355685
URI: http://eprints.soton.ac.uk/id/eprint/355685
ISSN: 1066-5099
PURE UUID: 69b09912-51af-4138-84d6-30ee82c8294d
ORCID for Franchesca D. Houghton: ORCID iD orcid.org/0000-0002-5167-1694

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Date deposited: 03 Sep 2013 15:16
Last modified: 09 Jan 2022 03:21

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Contributors

Author: Martin J. Pfeiffer
Author: Telma C. Esteves
Author: Sebastian T. Balbach
Author: Marcos J. Araúzo-Bravo
Author: Martin Stehling
Author: Anna Jauch
Author: Caroline Schwarzer
Author: Michele Boiani

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