Regenerated bovine fetal fibroblasts support high blastocyst development following nuclear transfer

Cloning
L LiuM E Westhusin

Abstract

Regenerated bovine fetal fibroblast cells were derived from a fetus cloned from an adult cow and passaged every 2-3 days. Serum starvation was performed by culturing cells in DMEM/F-12 supplemented with 0.5% FCS for 1-3 days. In vitro matured bovine oocytes were enucleated by removing the first polar body and a small portion of cytoplasm containing the metaphase II spindle. Cloned embryos were constructed by electrofusion of fetal fibroblast cells with enucleated bovine oocytes, electrically activated followed by 5 h culture in 10 microg/mL cycloheximide + 5 microg/mL cytochalasin B, and then cultured in a B2 + vero-cell co-culture system. A significantly higher proportion of fused embryos developed to blastocysts by day 7 when nuclei were exposed to oocyte cytoplasm prior to activation for 120 min (41.2%) compared to 0-30 min (28.2%, p < 0.01). Grade 1 blastocyst rates were 85.1% and 73.3%, respectively. The mean number of nuclei per grade 1 blastocyst was significantly greater for 120 min exposure (110.63 +/- 7.19) compared to 0-30 min exposure (98.67 +/- 7.94, p < 0.05). No significant differences were observed in both blastocyst development (37.4% and 30.6%) and mean number of nuclei per blastocyst (103.59 +/- 6.6 and 107.0...Continue Reading

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Citations

Jan 22, 2004·Cloning and Stem Cells·Cesare GalliGiovanna Lazzari
Feb 20, 2009·Cloning and Stem Cells·Mohammad Shamim HosseinWoo Suk Hwang
Nov 15, 2003·Reproductive Biology and Endocrinology : RB&E·X Cindy TianXiangzhong Yang
Dec 21, 2007·The Journal of Reproduction and Development·Tatsuya FujimuraHiroshi Nagashima
Jan 3, 2006·Reproduction : the Official Journal of the Society for the Study of Fertility·K I AstonK L White
Nov 17, 2010·Izvestiia Akademii nauk. Seriia biologicheskaia·G P MalenkoO I Stepanov
Mar 1, 2016·Frontiers in Microbiology·Patrycja A KobiereckaAgnieszka K Wyszyńska
Jan 7, 2016·Frontiers in Microbiology·Olaf TycPaolina Garbeva
Feb 22, 2013·Molecular Reproduction and Development·Simret BetshaDawit Tesfaye
Apr 12, 2011·Molecular Reproduction and Development·Michael C GoldingCharles R Long
Mar 11, 2004·Differentiation; Research in Biological Diversity·Keith E Latham
Apr 22, 2014·Cell Stem Cell·Young Gie ChungDong Ryul Lee

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