A new approach to cyropreservation of large equine embryos by vitrification after blastocoel micromanipulation.
J. Scherzer, R. A. Fayrer-Hosken, L. Ray and G. Heusner
Reproduction, Fertility and Development 19(1) 225 - 226 (abstracts from IETS 2007)SUMMARY (EXCERPT)
Vitrified large equine embryos >800 µm recovered on Day 8 after ovulation have not been successfully transferred yet. In this study, we examined the effects of reduction of the blastocoelic fluid and microinfusion of a cryopreservative prior to vitrification on pregnancy outcome.
To facilitate aspiration and microinfusion, a laser system (XYclone; Hamilton Thorne Biosciences, Beverly, MA, USA) was applied. Approximately 20% of the total blastocoelic fluid was removed using a standard micromanipulator and microcapillary system (Eppendorf). All embryos were then vitrified as previously described (Eldridge-Panuska et al. 2005 Theriogenology 63, 1308–1319).
None of the control embryos nor the split embryo in the treatment group led to the formation of an embryonic vesicle. However, the blastocyst, which had undergone both aspiration of blastocoelic fluid and microinfusion of VS1 (#5), had formed an embryonic vesicle at Day 15 after ovulation. During a further exam on Day 28, the uterine tone in the recipient was still increased, but ultrasonography revealed resorption of the embryo, which was probably caused by heat stress. Nevertheless, we will test this protocol for the cryopreservation of large equine embryos on a larger scale during the next breeding season.
Laser-assisted blastocyst dissection and subsequent cultivation of embryonic stem cells in a serum/cell free culture system: applications and preliminary results in a murine model.
Noriko Tanaka1, Takumi Takeuchi1, Queenie V Neri1, Eric Scott Sills2 and Gianpiero D Palermo1
1Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, New York, NY 10021, USA
2Department of Obstetrics, Gynecology and Reproductive Research, Murphy Medical Center, Murphy, NC, USA
Journal of Translational Medicine 2006, 4 : 20 doi:10.1186/1479-5876-4-20
Effects of Caffeine on the In Vitro Development of Bovine Nuclear Transfer Embryos.
Maalouf WE, Lee J-H, Campbell KHS. Animal Development and Biotechnology Group, School of Biosciences, University of Nottingham. Presented at IETS 2006
XYClone™ Laser System: A Novel Technique for Mouse Intracytoplasmic Sperm Injection (ICSI). William A. Richie, Embryologist, Division of Gene Expression and Development, Roslin Institute, Roslin Midlothian UK EH25 9PS and K. Bradley, Hamilton Thorne Biosciences, Beverly, MA 01915 USA
A New Method to Produce Mice Fully Derived from Embryonic Stem Cells. William T. Poueymirou, Wojtek Auerbach, Joseph Hickey, Jennifer Escaravage, David Frendewey, Niels C. Adams, George D. Yancopoulos, Thomad D. DeChiara, and David M. Valenzuela. Regerneron Pharmaceuticals, Inc. 777 Old Saw Mill River Road., tarrytown, NY, USA. Presented at the 2005 Transgenic Technology Meeting, Barcelona, Spain, September 11-13, 2005
A Feeder Cell- and Serum-Free System to Harvest Embryonic Stem Cells From Laser Dissected Blastocysts. T. Takeuchi, N. Tanaka, Q.V. Neri, Z. Rosenwaks, G.D. Palermo. Cornell University, New York, NY. Abstract from American Society of Reproductive Medicine, September 2005
Laser zona drilling does not induce hsp70i transcription in blastomeres of eight-cell mouse embryos.
Cristina Hartshorn, Ph.D., Aleksandra Anshelevich, B.S.,
Lawrence J. Wangh, Ph.D.
To assess whether zona drilling with a 1,480-nm laser
induces heat shock in eight-cell embryos, we measured hsp70i RNA levels
in sets of single blastomeres isolated after laser treatment of mouse
embryos that had or had not been heated at 43°C. Unlike heating, laser
zona drilling did not stimulate hsp70i expression, even in the
blastomeres closest to the laser beam, corroborating the safety of this
procedure for assisted reproduction.
(Fertil Steril 2005;84:1547–50. ©2005 by American Society for Reproductive Medicine.)
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