Effect of cell confluence on production of cloned mice using an inbred embryonic stem cell line

Gao, Shaorong, McGarry, Michelle, Ferrier, Tricia, Pallante, Benedetta, Gasparrini, Bianca, Fletcher, Judy, Harkness, Linda, De Sousa, Paul, McWhir, Jim and Wilmut, Ian (2003) Effect of cell confluence on production of cloned mice using an inbred embryonic stem cell line. Biology of Reproduction, 68 2: 595-603. doi:10.1095/biolreprod.102.005819


Author Gao, Shaorong
McGarry, Michelle
Ferrier, Tricia
Pallante, Benedetta
Gasparrini, Bianca
Fletcher, Judy
Harkness, Linda
De Sousa, Paul
McWhir, Jim
Wilmut, Ian
Title Effect of cell confluence on production of cloned mice using an inbred embryonic stem cell line
Journal name Biology of Reproduction   Check publisher's open access policy
ISSN 0006-3363
1529-7268
Publication date 2003-02-01
Year available 2002
Sub-type Article (original research)
DOI 10.1095/biolreprod.102.005819
Open Access Status Not yet assessed
Volume 68
Issue 2
Start page 595
End page 603
Total pages 9
Place of publication Cary, NC, United States
Publisher Oxford University Press
Language eng
Abstract Mice have been successfully cloned from both somatic cells and hybrid embryonic stem (ES) cells. Heterozygosity of the donor ES cell genome has been suggested as a crucial factor for long-term survival of cloned mice. In the present study, an inbred ES cell line, HM-1 (129/Ola), and a well-tested ES cell line, R1 (129/Sv × 129/Sv-CP), were used as donor cells to evaluate the developmental potential of nuclear transfer embryos. We found that ES cell confluence dramatically affects the developmental potential of reconstructed embryos. With the ES cell line HM-1 and 80-90% confluence, 49% of reconstructed embryos developed to the morula/blastocyst stage, 9% of these embryos developed to live pups when transferred to the surrogate mothers, and 5 of 18 live pups survived to adulthood. By contrast, at 60-70% confluence, only 22% of embryos developed to the morula/blastocyst stage, and after transfer, only a single fetus reached term. Consistent with previous reports, the nuclei of R1 ES cells were also shown to direct development to term, but no live pups were derived from cells at later passages (>20). Our results show that the developmental potential of reconstructed embryos is determined by both cell confluence and cell passage. These results also demonstrate that the inbred ES cell line, HM-1, can be used to produce viable cloned mice, although less efficiently than most heterozygous ES cell lines.
Keyword Developmental biology
Embryo
Oocyte development
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Wed, 23 Sep 2015, 16:21:22 EST by Linda Harkness on behalf of Learning and Research Services (UQ Library)