Human Reproduction, Vol. 11, No. 12, pp. 2703-2712, 1996
© 1996 European Society of Human Reproduction and Embryology
research-article |
Fertilization and early embryology: Alleviation of the ‘2-cell block’ and development to the blastocyst of CF1 mouse embryos: role of amino acids, EDTA and physical parameters
Laboratories of Human and Animal Reproductive Biology, Institute of Reproduction and Development, Monash University Victoria, Australia
Correspondence: 1To whom correspondence should be addressed at. Institute of Reproduction and Development, Monash Medical Centre, 246 Clayton Road, Clayton, Melbourne, Victoria 3168, Australia
The role of amino acids, ethylenediaminetetraacetic acid (EDTA), transferrin, oxygen, glucose, glutamine, taurine and ammonium in CF1 mouse zygote development in culture was examined. Non-essential amino acids and glutamine were shown to alleviate the 2-cell block in culture, and acted in synergy with EDTA to facilitate development to the blastocyst stage. In the presence of amino acids and EDTA, transferrin conferred no beneficial effect Development of zygotes was significantly impaired if amino acids were removed from the collection medium, even when they were subsequently cultured in the presence of amino acids. Zygote development to the blastocyst stage was significantly improved when modular incubator chambers were used compared to using a conventional incubator, and when an oxygen concentration of 7% was used as opposed to 20%. Addition of taurine to medium containing non-essential amino acids had no effect on embryo development, whereas the removal of glutamine and/or glucose from the culture medium significantly reduced blastocyst cell number. Removal of glucose from the culture medium also resulted in a significant decrease in implantations. Ammonium, generated from the breakdown of amino acids, significantly reduced blastocyst development EDTA was found to confer its beneficial effects during the first 48 h of culture, and indeed was inhibitory during the second 48 h, resulting in loss of subsequent viability. In summary, the data demonstrate that development of CF1 zygotes to the blastocyst stage is readily achievable. In the presence of non-essential amino acids and glutamine the removal of glucose is detrimental to CF1 mouse embryo development in culture and reduces subsequent viability. Optimal development and maintenance of viability requires more than one culture medium to support the preimplantation period.
Key words: ammonium/culture/EDTA/oxygen/viability
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  R. G. Sturmey, J. A. Hawkhead, E. A. Barker, and H. J. Leese DNA damage and metabolic activity in the preimplantation embryo Hum. Reprod., January 1, 2009; 24(1): 81 - 91. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D.H. Betts and P. Madan Permanent embryo arrest: molecular and cellular concepts Mol. Hum. Reprod., August 1, 2008; 14(8): 445 - 453. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Watkins, A. Wilkins, C. Cunningham, V. H. Perry, M. J. Seet, C. Osmond, J. J. Eckert, C. Torrens, F. R. A. Cagampang, J. Cleal, et al. Low protein diet fed exclusively during mouse oocyte maturation leads to behavioural and cardiovascular abnormalities in offspring J. Physiol., April 15, 2008; 586(8): 2231 - 2244. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Pantaleon, J. Scott, and P. L. Kaye Nutrient Sensing by the Early Mouse Embryo: Hexosamine Biosynthesis and Glucose Signaling During Preimplantation Development Biol Reprod, April 1, 2008; 78(4): 595 - 600. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. D. Biggers, L. McGinnis, and M. C. Summers Reply: One-step versus two-step culture of mouse preimplantation embryos Hum. Reprod., July 1, 2006; 21(7): 1936 - 1939. [Full Text] [PDF]
|
|
|
|
|
|
D. Feil, M. Lane, C. T. Roberts, R. L. Kelley, L. J. Edwards, J. G. Thompson, and K. L. Kind Effect of culturing mouse embryos under different oxygen concentrations on subsequent fetal and placental development J. Physiol., April 1, 2006; 572(1): 87 - 96. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Gopichandran and H. J Leese The effect of paracrine/autocrine interactions on the in vitro culture of bovine preimplantation embryos Reproduction, February 1, 2006; 131(2): 269 - 277. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. G Katz-Jaffe, D. W Linck, W. B Schoolcraft, and D. K Gardner A proteomic analysis of mammalian preimplantation embryonic development Reproduction, December 1, 2005; 130(6): 899 - 905. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.D. Biggers, L.K. McGinnis, and J.A. Lawitts One-step versus two-step culture of mouse preimplantation embryos: is there a difference? Hum. Reprod., December 1, 2005; 20(12): 3376 - 3384. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Hadi, M.-A. Hammer, C. Algire, T. Richards, and J. M. Baltz Similar Effects of Osmolarity, Glucose, and Phosphate on Cleavage past the 2-Cell Stage in Mouse Embryos from Outbred and F1 Hybrid Females Biol Reprod, January 1, 2005; 72(1): 179 - 187. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. M Orsi and H. J Leese Ammonium exposure and pyruvate affect the amino acid metabolism of bovine blastocysts in vitro Reproduction, January 1, 2004; 127(1): 131 - 140. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Emiliani, A. Delbaere, A.-S. Vannin, J. Biramane, M. Verdoodt, Y. Englert, and F. Devreker Similar delivery rates in a selected group of patients, for day 2 and day 5 embryos both cultured in sequential medium: a randomized study Hum. Reprod., October 1, 2003; 18(10): 2145 - 2150. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Lane and D. K. Gardner Ammonium Induces Aberrant Blastocyst Differentiation, Metabolism, pH Regulation, Gene Expression and Subsequently Alters Fetal Development in the Mouse Biol Reprod, October 1, 2003; 69(4): 1109 - 1117. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Van Thuan, H. Harayama, and M. Miyake Characteristics of Preimplantational Development of Porcine Parthenogenetic Diploids Relative to the Existence of Amino Acids In Vitro Biol Reprod, December 1, 2002; 67(6): 1688 - 1698. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Lane, J. M. Maybach, and D. K. Gardner Addition of ascorbate during cryopreservation stimulates subsequent embryo development Hum. Reprod., October 1, 2002; 17(10): 2686 - 2693. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Mortimer Human blastocyst development media Hum. Reprod., December 1, 2001; 16(12): 2725 - 2725. [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Rogers and E. S. Hunter III Redox Redux: A Closer Look at Conceptal Low Molecular Weight Thiols Toxicol. Sci., July 1, 2001; 62(1): 1 - 3. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
L. J. Van Winkle Amino Acid Transport Regulation and Early Embryo Development Biol Reprod, January 1, 2001; 64(1): 1 - 12. [Abstract] [Full Text]
|
|
|
|
|
|
M. C. Summers, L. K. McGinnis, J. A. Lawitts, M. Raffin, and J. D. Biggers IVF of mouse ova in a simplex optimized medium supplemented with amino acids Hum. Reprod., August 1, 2000; 15(8): 1791 - 1801. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. E. Neganova, G. G. Sekirina, and U. Eichenlaub-Ritter Surface-expressed E-cadherin, and mitochondrial and microtubule distribution in rescue of mouse embryos from 2-cell block by aggregation Mol. Hum. Reprod., May 1, 2000; 6(5): 454 - 464. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Tao and H. Niemann Cellular characterization of blastocysts derived from rabbit 4-, 8- and 16-cell embryos and isolated blastomeres cultured in vitro Hum. Reprod., April 1, 2000; 15(4): 881 - 889. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.H. McKiernan and B.D. Bavister Culture of one-cell hamster embryos with water soluble vitamins: pantothenate stimulates blastocyst production Hum. Reprod., January 1, 2000; 15(1): 157 - 164. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. K. Gardner, H. Rodriegez-Martinez, and M. Lane Fetal development after transfer is increased by replacing protein with the glycosaminoglycan hyaluronan for mouse embryo culture and transfer Hum. Reprod., October 1, 1999; 14(10): 2575 - 2580. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T.E. Steeves and D.K. Gardner Temporal and Differential Effects of Amino Acids on Bovine Embryo Development in Culture Biol Reprod, September 1, 1999; 61(3): 731 - 740. [Abstract] [Full Text]
|
|
|
|
|
|
M. Kolajova and J. M. Baltz Volume-Regulated Anion and Organic Osmolyte Channels in Mouse Zygotes Biol Reprod, April 1, 1999; 60(4): 964 - 972. [Abstract] [Full Text]
|
|
|
|
|
|
J. C.M. Dumoulin, C. J.J. Meijers, M. Bras, E. Coonen, J. P.M. Geraedts, and J. L.H. Evers Effect of oxygen concentration on human in-vitro fertilization and embryo culture Hum. Reprod., February 1, 1999; 14(2): 465 - 469. [Abstract] [Full Text] [PDF]
|
|