Journal article
Developmental capacity of ferret embryos by nuclear transfer using G0/G1-phase fetal fibroblasts
Biology of reproduction, Vol.68(6), pp.2297-2303
2003
DOI: 10.1095/biolreprod.102.012369
PMID: 12606336
Abstract
With the ultimate goal of establishing experimental protocols necessary for cloning ferrets, the present study has established parameters for the reconstruction of ferret embryos by nuclear transfer (NT) using G0/G1-phase donor fetal fibroblasts. Cumulus-oocyte complexes were harvested from superovulated ferrets and cultured in maturation medium for 24 h. Matured oocytes were then enucleated and injected with the fibroblast nuclei derived from 14–16-h serum-starved cells. Reconstructed embryos were then activated by a combination of electric pulses and chemical stimulations. Subsequently, the reconstructed and activated embryos were either cultured in vitro or transferred to pseudopregnant ferrets to evaluate their developmental capacity in vitro and in vivo. Our results demonstrated that 56.3% of reconstructed embryos (n = 187) cleaved, while 26.0% and 17.6% developed to morula and blastocyst phases in vitro, respectively. The blastocysts derived from NT embryos demonstrated normal morphology by differentially staining as compared to normal blastocysts developed in vivo following fertilization. In vivo developmental studies at 21 days posttransplantation demonstrated 8.8% of reconstructed embryos (n = 91) implanted into the uterine lining of recipients, while 3.3% formed fetuses. However, reconstructed embryos (n = 387) failed to develop to term (42 days). These results demonstrate donor nuclei of G0/G1-phase fetal fibroblast cells can be reprogrammed to support the development of reconstructed ferret embryos in vitro and in vivo; however, a significant third-trimester block occurs preventing full-term development.
Details
- Title: Subtitle
- Developmental capacity of ferret embryos by nuclear transfer using G0/G1-phase fetal fibroblasts
- Creators
- Ziyi Li - Department of Anatomy & Cell Biology, College of Medicine, University of Iowa, Iowa City, Iowa 52242, United StatesMaryam Rezaei Sabet - Department of Anatomy & Cell Biology, College of Medicine, University of Iowa, Iowa City, Iowa 52242, United StatesQi Zhou - Unite de Biologie du Developpement et Biotechnologie, Institut National de la Recherche Agronomique, 78352 Jouy-en-Josas, FranceXiaoming Liu - Department of Anatomy & Cell Biology, College of Medicine, University of Iowa, Iowa City, Iowa 52242, United StatesWei Ding - Department of Anatomy & Cell Biology, College of Medicine, University of Iowa, Iowa City, Iowa 52242, United StatesYulong Zhang - Department of Anatomy & Cell Biology, College of Medicine, University of Iowa, Iowa City, Iowa 52242, United StatesJean-Paul RENARD - Unite de Biologie du Developpement et Biotechnologie, Institut National de la Recherche Agronomique, 78352 Jouy-en-Josas, FranceJohn F ENGELHARDT - Department of Anatomy & Cell Biology, College of Medicine, University of Iowa, Iowa City, Iowa 52242, United States
- Resource Type
- Journal article
- Publication Details
- Biology of reproduction, Vol.68(6), pp.2297-2303
- DOI
- 10.1095/biolreprod.102.012369
- PMID
- 12606336
- NLM abbreviation
- Biol Reprod
- ISSN
- 0006-3363
- eISSN
- 1529-7268
- Publisher
- Society for the Study of Reproduction; Madison, WI
- Language
- English
- Date published
- 2003
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984025305002771
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