Abstract
Most embryo culture media are still supplemented with proteins rather than with nonprotein macromolecules or recombinant protein products. HSA is probably the most common supplement followed by globulin-enriched preparations. Serum supplementation and Co-Culture of embryos belong to the past. Defined nonprotein or recombinant protein supplements are becoming a viable alternative during gamete and embryo manipulation procedures. Biological protein supplements are still preferred for any extended period of embryo culture. Understanding the goals and purpose of supplemented macromolecules in embryo culture media during each step of the laboratory IVF process should assist us in choosing the safest and most consistent macromolecule for each step, but also selecting a product that has the capability of delivering the best clinical outcome. Each batch of biological protein supplement is unique, even if supplied by the same manufacturer. Each lot of protein supplement typically contains many lot-specific, potentially harmful, and unintended hormone and protein contaminants. Macromolecular embryo culture medium supplements should be identified as one of the highest risk factors in an IVF laboratory that may contribute towards clinical compromise. All efforts should be made to use a proven batch of supplement for as long as the expiration date will allow. The beneficial effect of more complex protein supplements is evident after the activation of the embryonic genome and probably due to the presence of growth factors. Lower live-birth rates due to suboptimum protein supplementation may be a direct result of the preferential loss of female embryos. When deciding on a culture system, thought should be given specifically to the interaction between the culture medium and the macromolecular supplement. Ready-to-use pre-supplemented culture media may be advisable over a more complex product if a comprehensive macromolecular quality management program is not feasible. However, the question remains as to whether the increasing simplification of embryo culture media supplements is ready for large-scale clinical use.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Steptoe PC, Edwards RG (1978) Birth after the reimplantation of a human embryo. Lancet 2(8085):366
Rock J, Menkin MF (1944) In vitro fertilization and cleavage of human ovarian eggs. Science 100(2588):105–107
McLaughlin L (1982) The Pill, John Rock and the Church. A biography of a revolution, 1st edn. Little, Brown and Company, Boston
Steptoe PC, Edwards RG, Purdy JM (1971) Human blastocysts grown in culture. Nature 229(5280):132–133
Dokras A, Sargent IL, Redman CWG (1993) Sera from woman with unexplained infertility inhibit both mouse and human embryo growth in vitro. Fertil Steril 60:285–292
Kemeter P, Feightinger W (1984) Pregnancy following in vitro fertilization and embryo transfer using pure human serum as culture and transfer medium. Fertil Steril 41(6):936–937
Staessen C, Van Den Abbeel E, Carle M et al (1990) Comparison between human serum and Albuminar-20™ supplement for in vitro fertilization. Hum Reprod 5(3):336–341
Leveille MC, Carnegie J, Tanphaichitr N (1992) Effects of human sera and human serum albumin on mouse embryo culture. J Assist Reprod Genet 9(1):45–52
Weathersbee PS, Pool TB, Ord T (1995) Synthetic serum substitute (SSS): a globulin-enriched protein supplement for human embryo culture. J Assist Reprod Genet 12(6):354–360
Pool TB (2004) Development of culture media for human assisted reproductive technology. Fertil Steril 81:287–289
Alberda AT, Van Os HC, Zeilmaker GH (1989) Hepatitis B virus infectie bij vrouwen behandeld met in vitro fertilisatie. Neederlands Tijdskrift Voor Geneeskunde 133:20–25
Klein R, Dumble LJ (1993) Transmission of Creutzfeldt-Jakob disease by blood transfusion. Lancet 341:768
Otani T, McDonough PG (1995) Earthquakes and prions. Fertil Steril 63:1137–1139
Wiemer KE, Cohen J, Amborskir GF et al (1989) In-vitro development and implantation of human embryos following culture on fetal bovine uterine fibroblast cells. Hum Reprod 4(5):595–600
Bongso A, Ng SC, Ratnam S (1990) Co-cultures: their relevance to assisted reproduction. Hum Reprod 5(8):893–900
Wiemer KE, Hoffman DI, Maxson WS et al (1993) Embryonic morphology and rate of implantation of human embryos following co-culture on bovine oviductal epithelial cells. Hum Reprod 8(1):97–101
Ménézo YJR, Sakkas D, Janny L (1995) Co-culture of the early human embryo: factors affecting human blastocyst formation in vitro. Microsc Res Tech 52(1):50–56
Rieger D, Grisart B, Semple E et al (1995) Comparison of the effects of oviductal cell co-culture and oviductal cell-conditioned medium on the development and metabolic activity of cattle embryos. J Reprod Fertil 105:91–98
Forsdahl F, Bertheussen K, Bungum LJ et al (1994) A study on extended culture time with embryo replacement at the morula and blastocyst stage. Hum Reprod 9:142–143
Barnes FL, Crombie A, Gardner DK et al (1995) Blastocyst development and pregnancy after in vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching. Hum Reprod 10:3243–3247
Behr B, Pool TB, Milki A et al (1999) Preliminary clinical experience with human blastocyst development in vitro without co-culture. Hum Reprod 14(2):454–457
Dugan KJ, Shalika S, Smith RD et al (1997) Comparison of synthetic serum substitute and fetal cord serum as media supplements for in vitro fertilization: a prospective, randomized study. Fertil Steril 67(1):166–168
Laverge H, De Sutter P, Desmet R, et al (1997) Prospective randomized study comparing human serum albumin with fetal cord serum as protein supplement in culture medium for in-vitro fertilization (IVF). Hum Reprod 12 (10):2263–2266
Tucker KE, Hurst BS, Guadagnoli S et al (1996) Evaluation of synthetic serum substitute versus serum as protein supplementation for mouse and human embryo culture. J Assist Reprod Genet 13(1):32–37
Blake D, Svalander P, Jin M et al (2002) Protein supplementation of human IVF culture media. J Assist Reprod Genet 19(3):137–143
Bungum M, Humaidan P, Bungum L (2002) Recombinant human albumin as protein source in culture media used for IVF: a prospective randomized study. Reprod Biomed Online 4(3):233–236
Takenaka M, Horiuchi T (2007) Recombinant human albumin supports mouse blastocyst development and improves fetal development. Reprod Med Biol 6(4):195–201
Gardner DK, Lane M (1998) Culture of viable human blastocysts in defined sequential serum-free media. Hum Reprod 13(Suppl 3):148–159
Ali J, Shahata MA, Al-Natsha SD (2000) Formulation of a protein-free medium for human assisted reproduction. Hum Reprod 15(1):145–156
Friedler S, Schachter M, Strassburger D et al (2007) A randomized clinical trial comparing recombinant hyaluronan/recombinant albumin versus human tubal fluid for cleavage stage embryo transfer in patients with multiple IVF-embryo transfer failure. Hum Reprod 22(9):2444–2448
Simon A, Safran A, Revel A et al (2003) hyaluronic acid can successfully replace albumin as the sole macromolecule in a human embryo transfer medium. Fertil Steril 79(6):1434–1438
Tanikawa T, Harada T, Ito M et al (1999) Globulins in protein supplements promote the development of preimplantation embryos. J Assist Reprod Genet 16:555–557
Meintjes M, Chantilis SJ, Ward DC et al (2009) A randomized controlled study of human serum albumin and serum substitute supplement as protein supplements for IVF culture and the effect on live birth rates. Hum Reprod 24(4):782–789
Richter KS (2008) The importance of growth factors for preimplantation embryo development and in vitro culture. Curr Opin Obstet Gynecol 20(3):292–304
Behboodi E, Anderson GB, BonDurant RH et al (1994) Birth of large calves that developed from in-vitro derived bovine embryos. Theriogenology 44:227–232
Hardy K, Spanos S (2002) Growth factor expression and function in the human and mouse preimplantation embryo. J Endocrinol 172:221–236
Lane M, Hooper K, Gardner DK (2001) Effect of essential amino acids on mouse embryo viability and ammonium production. J Assist Reprod Genet 18:519–525
Alvarez JG, Storey BT (1995) Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa. Mol Reprod Dev 42:334–346
Hambiliki F, Ljunger E, Olof Karlström P et al (2009) Hyaluronan-enriched transfer medium in cleavage-stage frozen-thawed embryo transfers increases implantation rate without improvement of delivery rate. Fertil Steril 94(5):1669–1673
Graham MC, Partridge AB, Lewis V et al (1995) A prospective comparison of synthetic serum substitute and human serum albumin in culture for in vitro fertilization-embryo transfer. Fertil Steril 64(5):1036–1038
Rienzi L, Ubaldi F, Anniballo R et al (1998) Preincubation of human oocytes may improve fertilization and embryo quality after intracytoplasmic sperm injection. Hum Reprod 13(4):1014–1019
Ho JY, Chen MJ, Yi YC et al (2003) The effect of preincubation period of oocytes on nuclear maturity, fertilization rate, embryo quality and pregnancy outcome in IVF and ICSI. J Assist Reprod Genet 20(9):358–364
Guerif F, Cadoret V, Poindron J et al (2003) Overnight incubation improves selection of frozen-thawed blastocysts for transfer: preliminary study using supernumary embryos. Theriogenology 60(8):1457–1466
Gorrill MJ, Rinehart JS, Tamhane AC et al (1991) Comparison of the hamster sperm motility assay to the mouse one-cell and two-cell embryo bioassays as quality control tests for in vitro fertilization. Fertil Steril 55(2):345–354
Claasens OE, Harrison KL (1999) Optimizing sensitivity of human sperm motility assay for embryology toxicity testing. In: Proceedings of the 11 th world congress on IVF and human reproductive genetics. Sydney, Australia
Pemble LB, Kaye PL (1976) Whole protein uptake and metabolism by mouse blastocysts. J Reprod Fertil 78:149–157
O’Neil C (2005) The role of PAF in embryo physiology. Hum Reprod Update 11(3):215–228
Ammit AJ, O’Neill C (1997) Studies of the nature of the binding by albumin of platelet-activating factor released from cells. J Biol Chem 272:18772–18778
O’Neil C (1997) Evidence for the requirement of autocrine growth factors for development of mouse preimplantation embryos in vitro. Biol Reprod 56:229–237
Paria BC, Dey SK (1990) Preimplantation embryo development in vitro: cooperative interactions among embryos and role of growth factors. Proc Natl Acad Sci USA 87(12):4756–4760
Hink JH, Hidalgo J, Seeberg VP et al (1957) Preparation and properties of a heat-treated human plasma protein fraction. Vox Sanguinis 2(3):174–186
Ben-Yosef D, Yovel I, Schwartz T et al (2001) Increasing synthetic serum substitute (SSS) concentrations in P1 glucose/phosphate-free medium improves implantation rate: a comparative study. J Assist Reprod Genet 18(11):588–592
Mosley AK, Brouwer KL (1997) Heat treatment of human serum to inactivate HIV does not alter protein binding of selected drugs. Ther Drug Monit 19:477–479
Snyman E, Van der Merwe JV (1986) Endotoxin-polluted medium in a human in vitro fertilization program. J In Vitro Fertil Embryo Transfer 5:335–342
Mahani IM, Davar R (2007) Hyaluronic acid versus albumin in human embryo transfer medium. East Mediterr Health J 13(4):876–880
Gray CW, Morgan PM, Kane MT (1992) Purification of an embryotrophic factor from commercial bovine serum albumin and its identification s citrate. J Reprod Fertil 94:471–480
Fishel S, Jackson P, Webster J et al (1988) Endotoxins in culture medium for human in vitro fertilization. Fertil Steril 49:108–111
Nagata Y, Shirakawa K (1996) Setting standards for the levels of endotoxin in the embryo culture media of human in vitro fertilization and embryo transfer. Fertil Steril 65:614–619
Kimura K, Spate LD, Green MP et al (2005) Effects of d-glucose concentration, d-fructose, and inhibitors of enzymes of the pentose phosphate pathway on the development and sex ratio of bovine blastocysts. Mol Reprod Dev 72:201–207
Chang HJ, Lee JR, Jee BC et al (2009) Impact of blastocyst transfer on offspring sex ratio and the monozygotic twinning rate: a systematic review and meta-analysis. Fertil Steril 91(6):2381–2390
Bermejo-Alvarez P, Rizos D, Lonergan P et al (2011) Transcriptional sexual dimorphism during preimplantation embryo development and its consequences for developmental competence and adult health and disease. Reproduction 141:563–570
Gutierrez-Adan A, Perez-Crispo M, Fernandez-Gonzalez R et al (2006) Developmental consequences of sexual dimorphism during preimplantation embryonic development. Reprod Domest Anim 41(Suppl 2):54–62
Holm P, Walker SK, Seamark RF (1996) Embryo viability, duration of gestation and birth weight in sheep after transfer of in vitro matured and in vitro fertilized zygotes cultured in vitro or in vivo. J Reprod Fertil 107:175–181
Pool TB, Martin JE (1994) High continuing pregnancy rates after in vitro fertilization-embryo transfer using medium supplemented with a plasma protein fraction containing α and β globulins. Fertil Steril 61:714–719
Acknowledgments
I thank the dedicated physicians, embryologists, nurses, and administrative personnel working in our program who are always willing to support new studies and investigational endeavors. I gratefully acknowledge Vitrolife Inc. (Inglewood, Colorado) for their sponsorship and technical support of several investigations and studies in our quest to continually improve culture conditions and embryo culture medium formulations.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media New York
About this protocol
Cite this protocol
Meintjes, M. (2012). Media Composition: Macromolecules and Embryo Growth. 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_8
Download citation
DOI: https://doi.org/10.1007/978-1-61779-971-6_8
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-970-9
Online ISBN: 978-1-61779-971-6
eBook Packages: Springer Protocols