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Summary. One-cell CF-1 x B6SJLF1/J embryos, which usually exhibit a 2-cell block to development in vitro, have been cultured to the blastocyst stage using CZB medium and a glucose washing procedure. CZB medium is a further modification of modified BMOC-2 containing an increased lactate/pyruvate ratio of 116, 1 mM-glutamine and 0·1 mM-EDTA but lacking glucose. Continuous culture of one-cell embryos in CZB medium allowed 83% of embryos to develop beyond the 2-cell stage of which 63% were morulae at 72 h of culture, but blastocysts did not develop. However, washing embryos into CZB medium containing glucose after 48 h of culture (3-4-cell stage) was sufficient to allow development to proceed, with 48% of embryos reaching the blastocyst stage by 96 h of culture. Exposure of embryos to glucose was only necessary from the 3–4-cell stage through the early morula stage since washing back into medium CZB without glucose at 72 h of culture still promoted the development of 50% of embryos to the blastocyst stage. The presence of glucose in this medium for the first 48 h of culture (1-cell to 4-cell stage) was detrimental to embryo development. Glutamine, however, exerted a beneficial effect on embryo development from the 1-cell to the 4-cell stage although its presence was not required for development to proceed during the final 48 h of culture. Blastocysts which developed under optimum conditions contained an average of 33·7 total cells. The in-vitro development of 1-cell embryos beyond the 2-cell stage in response to the removal of glucose and the addition of glutamine to the culture medium suggests that glucose may block some essential metabolic process, and that glutamine may be a preferred energy substrate during early development for these mouse embryos.
Keywords: mouse; embryo; 2-cell block; glutamine; glucose
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M. A. Szczygiel, H. Kusakabe, R. Yanagimachi, and D. G. Whittingham Intracytoplasmic Sperm Injection Is More Efficient than In Vitro Fertilization for Generating Mouse Embryos from Cryopreserved Spermatozoa Biol Reprod, October 1, 2002; 67(4): 1278 - 1284. [Abstract] [Full Text] [PDF]
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B. A. McAvey, G. B. Wortzman, C. J. Williams, and J. P. Evans Involvement of Calcium Signaling and the Actin Cytoskeleton in the Membrane Block to Polyspermy in Mouse Eggs Biol Reprod, October 1, 2002; 67(4): 1342 - 1352. [Abstract] [Full Text] [PDF]
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K. Mizuno, K. Hoshi, and T. Huang Fertilization and embryo development in a mouse ICSI model using human and mouse sperm after immobilization in polyvinylpyrrolidone Hum. Reprod., September 1, 2002; 17(9): 2350 - 2355. [Abstract] [Full Text] [PDF]
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S. Gao, B. Gasparrini, M. McGarry, T. Ferrier, J. Fletcher, L. Harkness, P. De Sousa, and I. Wilmut Germinal Vesicle Material Is Essential for Nucleus Remodeling after Nuclear Transfer Biol Reprod, September 1, 2002; 67(3): 928 - 934. [Abstract] [Full Text] [PDF]
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R. L. Brown, T. Ord, S. B. Moss, and C. J. Williams A-Kinase Anchor Proteins as Potential Regulators of Protein Kinase A Function in Oocytes Biol Reprod, September 1, 2002; 67(3): 981 - 987. [Abstract] [Full Text] [PDF]
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Y. G. Chung, M. R.W. Mann, M. S. Bartolomei, and K. E. Latham Nuclear-Cytoplasmic "Tug of War" During Cloning: Effects of Somatic Cell Nuclei on Culture Medium Preferences of Preimplantation Cloned Mouse Embryos Biol Reprod, April 1, 2002; 66(4): 1178 - 1184. [Abstract] [Full Text] [PDF]
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T. E. Ludwig, J. M. Squirrell, A. C. Palmenberg, and B. D. Bavister Relationship Between Development, Metabolism, and Mitochondrial Organization in 2-Cell Hamster Embryos in the Presence of Low Levels of Phosphate Biol Reprod, December 1, 2001; 65(6): 1648 - 1654. [Abstract] [Full Text] [PDF]
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G. Manandhar and K. Toshimori Exposure of Sperm Head Equatorin after Acrosome Reaction and Its Fate after Fertilization in Mice Biol Reprod, November 1, 2001; 65(5): 1425 - 1436. [Abstract] [Full Text] [PDF]
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J.-S. Xu, T.-M. Cheung, S. Ting-Hon Chan, P.-C. Ho, and W. Shu-Biu Yeung Temporal Effect of Human Oviductal Cell and Its Derived Embryotrophic Factors on Mouse Embryo Development Biol Reprod, November 1, 2001; 65(5): 1481 - 1488. [Abstract] [Full Text] [PDF]
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J. Cozzi, F. Monier-Gavelle, N. Lievre, M. Bomsel, and J.P. Wolf Mouse Offspring after Microinjection of Heated Spermatozoa Biol Reprod, November 1, 2001; 65(5): 1518 - 1521. [Abstract] [Full Text] [PDF]
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D.-i. Fuchimoto, A. Mizukoshi, R. M. Schultz, S. Sakai, and F. Aoki Posttranscriptional Regulation of Cyclin A1 and Cyclin A2 During Mouse Oocyte Meiotic Maturation and Preimplantation Development Biol Reprod, October 1, 2001; 65(4): 986 - 993. [Abstract] [Full Text] [PDF]
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S. M. Hawes, Y. Gie Chung, and K. E. Latham Genetic and Epigenetic Factors Affecting Blastomere Fragmentation in Two-Cell Stage Mouse Embryos Biol Reprod, October 1, 2001; 65(4): 1050 - 1056. [Abstract] [Full Text] [PDF]
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J. Yu, N. B. Hecht, and R. M. Schultz Expression of MSY2 in Mouse Oocytes and Preimplantation Embryos Biol Reprod, October 1, 2001; 65(4): 1260 - 1270. [Abstract] [Full Text] [PDF]
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A. Ziyyat and A. Lefevre Differential gene expression in pre-implantation embryos from mouse oocytes injected with round spermatids or spermatozoa Hum. Reprod., July 1, 2001; 16(7): 1449 - 1456. [Abstract] [Full Text] [PDF]
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G. Leppens-Luisier, F. Urner, and D. Sakkas Facilitated glucose transporters play a crucial role throughout mouse preimplantation embryo development Hum. Reprod., June 1, 2001; 16(6): 1229 - 1236. [Abstract] [Full Text] [PDF]
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T. E. Ludwig, M. Lane, and B. D. Bavister Differential Effect of Hexoses on Hamster Embryo Development in Culture Biol Reprod, May 1, 2001; 64(5): 1366 - 1374. [Abstract] [Full Text]
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F. Devreker, K. Hardy, M. Van den Bergh, A.S. Vannin, S. Emiliani, and Y. Englert Amino acids promote human blastocyst development in vitro Hum. Reprod., April 1, 2001; 16(4): 749 - 756. [Abstract] [Full Text] [PDF]
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J. D. Biggers and L. K. McGinnis Evidence that glucose is not always an inhibitor of mouse preimplantation development in vitro Hum. Reprod., January 1, 2001; 16(1): 153 - 163. [Abstract] [Full Text] [PDF]
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