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Culture Systems: Single Step

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Embryo Culture

Part of the book series: Methods in Molecular Biology ((MIMB,volume 912))

Abstract

Culture media to support development of zygotes to the blastocyst stage is based either on a single medium or sequential (two-step) media. Single medium culture either with or without day 3 renewal is associated with simplified laboratory protocols and lower costs compared with sequential medium. There are currently insufficient clinical data to conclude that one system, either single or sequential, is superior to the other with regard to clinical performance. This chapter summarizes the rationale for use of a single medium, along with supporting animal and human data for culturing embryos in a single medium, renewed or not renewed.

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References

  1. Pauerstein CJ, Eddy CA (1979) The role of the oviduct in reproduction; our knowledge and our ignorance. J Reprod Fertil 55:223–229

    Article  PubMed  CAS  Google Scholar 

  2. Diaz S, Ortiz ME, Croxatto HB (1980) Studies on the duration of ovum transport by the human oviduct. III. Time interval between the luteinizing hormone peak and recovery of ova by transcervical flushing of the uterus in normal women. Am J Obstet Gynecol 137:116–121

    PubMed  CAS  Google Scholar 

  3. Lyons RA, Saridogan E, Djahanbakhch O (2006) The reproductive significance of human fallopian tube cilia. Hum Reprod Update 12:363–372

    Article  PubMed  CAS  Google Scholar 

  4. Biggers JD (1998) Reflection of the culture of preimplantation embryo. Int J Dev Biol 42:879–884

    PubMed  CAS  Google Scholar 

  5. Gardner DK, Lane M (1996) Alleviation of the ‘2-cell block’ and development to the blastocyst of CF1 mouse embryos: role of amino acids, EDTA and physical parameters. Hum Reprod 11:2703–2712

    Article  PubMed  CAS  Google Scholar 

  6. Leese HJ, Tay JI, Reischl J, Downing SJ (2001) Formation of fallopian tubal fluid: role of a neglected epithelium. Reproduction 121:339–346

    Article  PubMed  CAS  Google Scholar 

  7. Lane M, Gardner DK (2007) Embryo culture medium: which is the best? Best Pract Res Clin Obstet Gynaecol 21(1):83–100

    Article  PubMed  Google Scholar 

  8. Biggers JD (2000) Thoughts on embryo culture conditions. Reprod BioMed Online 4(1):30–38

    Google Scholar 

  9. Leese HJ (1991) Metabolism of the preimplantation mammalian embryo. Oxford Rev Reprod Biol 13:35–72

    CAS  Google Scholar 

  10. Leese HJ (1998) Human embryo culture: back to nature. J Assist Reprod Gene 15:466–468

    Article  CAS  Google Scholar 

  11. Biggers JD, Racowsky C (2002) The development of fertilized human ova to the blastocyst stage in KSOMAA medium: is a two-step protocol necessary? Reprod Biomed Online 5(2):133–140

    Article  PubMed  Google Scholar 

  12. Whitten WK (1956) Culture of tubal ova. Nature 177:96

    Article  PubMed  CAS  Google Scholar 

  13. McLaren A, Biggers JD (1958) Successful development and birth of mice cultivated in vitro as early as early embryos. Nature 182(4639):877–878

    Article  PubMed  CAS  Google Scholar 

  14. Whitten WK (1957) Culture of tubal ova. Nature 179:1081–1082

    Article  PubMed  CAS  Google Scholar 

  15. Biggers JD, Gwatkin RB, Brinster RL (1962) Development of mouse embryos in organ cultures of fallopian tubes on a chemically defined medium. Nature 194:747–749

    Article  PubMed  CAS  Google Scholar 

  16. Brinster RL (1972) Cultivation of the mammalian embryo. In: Rothblat G, Cristofalo V (eds) Growth, nutrition and metabolism of cells in culture. Academic, New York

    Google Scholar 

  17. Whittingham DG (1971) Culture of mouse ova. J Reprod Fertil 14:7–21

    CAS  Google Scholar 

  18. Chatot CL, Ziomek CA, Bavister BD, Lewis JL, Torres I (1989) An improved culture medium supports development of Random-bred 1-cell mouse embryos in vitro. J Reprod Fertil 86:679–688

    Article  PubMed  CAS  Google Scholar 

  19. Schini SA, Bavister BD (1988) Two-cell block to development of cultured hamster embryos is caused by phosphate and glucose. Biol Reprod 39:1183–1192

    Article  PubMed  CAS  Google Scholar 

  20. Thompson JG, Simpson AC, Pugh PA, Tervit HR (1992) Requirement for glucose during in vitro culture of sheep preimplantation embryos. Mol Reprod Dev 31:253–257

    Article  PubMed  CAS  Google Scholar 

  21. Takahashi Y, First NL (1992) In vitro development of bovine one-cell embryos: influence of glucose, lactate, pyruvate, amino acids and vitamins. Theriogenology 37:963–978

    Article  PubMed  CAS  Google Scholar 

  22. Quinn P (1995) Enhanced results in mouse and human embryo culture using a modified human tubal fluid medium lacking glucose and phosphate. J Assist Reprod Genet 12:97–105

    Article  PubMed  CAS  Google Scholar 

  23. Biggers JD, McGinnis L (2001) Evidence that glucose is not always an inhibitor of mouse preimplantation development in vitro. Hum Reprod 16(1):153–163

    Article  PubMed  CAS  Google Scholar 

  24. Miller JF, Schulz GA (1987) Amino acid content of preimplantation rabbit embryos and fluids of the reproductive tract. Biol Reprod 36:125–129

    Article  PubMed  CAS  Google Scholar 

  25. Gardner DK, Lane M (1993) Amino acids and ammonium regulate mouse embryo development in culture. Biol Reprod 48:377–385

    Article  PubMed  CAS  Google Scholar 

  26. Lane M, Gardner DK (2003) Ammonium induces aberrant blastocyst differentiation, metabolism, pH regulation, gene expression and subsequently alters fetal development in the mouse. Biol Reprod 69(4):1109–1117

    Article  PubMed  CAS  Google Scholar 

  27. Orsi NM, Leese HJ (2004) Ammonium exposure and pyruvate affect the amino acid metabolism of bovine blastocysts in vitro. Reproduction 127(1):131–140

    Article  PubMed  CAS  Google Scholar 

  28. Zander DL, Thompson JG, Lane M (2006) Perturbations in mouse embryo development and viability caused by ammonium are more severe after exposure at the cleavage stages. Biol Reprod 74(2):288–294

    Article  PubMed  CAS  Google Scholar 

  29. Virant-Klun I, Tomazevic T, Vrtacnik-Bokal E, Vogler A, Krsnik M, Meden-Vrtovec H (2006) Increased ammonium in culture medium reduces the development of human embryos to the blastocyst stage. Fertil Steril 85:526–528

    Article  PubMed  CAS  Google Scholar 

  30. Erbach GT, Bhatnagar P, Baltz JM, Biggers JD (1995) Zinc is a possible toxic contaminant of silicone oil in microdrop cultures of preimplantation mouse embryos. Hum Reprod 10:3248–3254

    PubMed  CAS  Google Scholar 

  31. Nasr-Esfahani MH (1992) Effects of glucose, glutamine, ethylenediaminetetraacetic acid and oxygen tension on the concentration of reactive oxygen species and on development of the mouse preimplantation embryo in vitro. J Reprod Fertil 96:219–231

    Article  PubMed  CAS  Google Scholar 

  32. Gardner DK, Lane M, Schoolcraft WB (2002) Physiology and culture of the human blastocyst. J Reprod Immunol 55(1–2):85–100

    Article  PubMed  CAS  Google Scholar 

  33. Summers MC, Biggers JD (2003) Chemically defined media and the culture of mammalian preimplantation embryos: historical perspective and current issues. Hum Reprod Update 9(6):557–82

    Article  PubMed  CAS  Google Scholar 

  34. Biggers JD, Summers MC (2008) Choosing a culture medium: making informed choices. Fertil Steril 90(3):473–483

    Article  PubMed  Google Scholar 

  35. Biggers JD, McGinnis LK, Lawitts JA (2005) One-step versus two-step culture of mouse preimplantation embryos: is there a difference? Hum Reprod 20(12):3376–3384

    Article  PubMed  CAS  Google Scholar 

  36. Perin PM, Maluf M, Nicolosi Foltran Januário DA, Nascimento Saldiva PH (2008) Comparison of the efficacy of two commercially available media for culturing one-cell embryos in the in vitro fertilization mouse model. Fertil Steril 90:1503–1510

    Article  PubMed  Google Scholar 

  37. Macklon NS, Pieters MH, Hassan MA, Jeucken PH, Eijkemans MJ, Fauser BC (2002) A prospective randomized comparison of sequential versus monoculture systems for in-vitro human blastocyst development. Hum Reprod 17(10):2700–2705

    Article  PubMed  CAS  Google Scholar 

  38. Reed ML, Hamic A, Thompson DJ, Caperton CL (2009) Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study. Fertil Steril 92(5):1783–1786

    Article  PubMed  Google Scholar 

  39. Sepulveda S, Garcia J, Arriaga E, Diaz J, Noriega-Portella L, Noriega-Hoces L (2009) In vitro development and pregnancy outcomes for human embryos cultured in either a single medium or in a sequential media system. Fertil Steril 91:1765–1770

    Article  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Obstetrics, Gynecology & Reproductive Biology, Brigham and Women’s Hospital, Boston, MA, USA

    Ronit Machtinger & Catherine Racowsky

Authors
  1. Ronit Machtinger
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  2. Catherine Racowsky
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Corresponding author

Correspondence to Catherine Racowsky .

Editor information

Editors and Affiliations

  1. , OB/GYN, University of Michigan, E. Catherine Street 1137, Ann Arbor, 48109-5622, Michigan, USA

    Gary D. Smith

  2. , Center for Reproductive Medicine, University of Michigan, Market Street 475, Ann Arbor, 48108, Michigan, USA

    Jason E. Swain

  3. Fertility Center of San Antonio, Medical Drive 4499, San Antonio, 78229, Texas, USA

    Thomas B. Pool

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© 2012 Springer Science+Business Media New York

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Machtinger, R., Racowsky, C. (2012). Culture Systems: Single Step. In: Smith, G., Swain, J., Pool, T. (eds) Embryo Culture. Methods in Molecular Biology, vol 912. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-971-6_12

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  • DOI: https://doi.org/10.1007/978-1-61779-971-6_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-970-9

  • Online ISBN: 978-1-61779-971-6

  • eBook Packages: Springer Protocols

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