Skip to main content Accessibility help
×
Hostname: page-component-848d4c4894-cjp7w Total loading time: 0 Render date: 2024-06-24T17:54:29.402Z Has data issue: false hasContentIssue false

Chapter 5 - Culture Media and Embryo Culture

Published online by Cambridge University Press:  15 April 2021

Kersti Lundin
Affiliation:
Sahlgrenska University Hospital, Gothenburg
Aisling Ahlström
Affiliation:
Livio Fertility Center, Gothenburg
Get access

Summary

Before considering the basis of embryo culture medium, it is worthwhile reflecting on its function. A detailed overview of embryo metabolism is given in Chapter 4; however, in brief, the embryo must satisfy changing demands for energy by consumption of nutrients from the external milieu (Lewis & Sturmey, 2015). In an in vivo setting, these needs are catered for in a dynamic manner by the secretions of the oviduct; in an in vitro situation, these requirements must be satisfied by the embryo culture medium. In addition to the provision of energy substrates, the medium must also satisfy basic physicochemical requirements. Primarily, the medium must facilitate buffering of pH in response both to changing environments to which the embryo is exposed and to excretion of metabolic waste products, notably lactic acid, which is released by cells with accompanying protons, causing pH to fall. Moreover, the culture medium must avoid inducing osmotic stress. One of the major consumers of cellular energy is the maintenance of intracellular ion composition, maintained through the action of ion pumps. Providing suitable osmolarity and pH are among the most basic requirements of any embryo culture medium.

Type
Chapter
Information
Publisher: Cambridge University Press
Print publication year: 2021

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aguilar, J, Reyley, M. The uterine tubal fluid : secretion, composition and biological effects. Anim Reprod. 2005;2:91105.Google Scholar
Almagor, M, Bejar, C, Kafka, I, Yaffe, H. Pregnancy rates after communal growth of preimplantation human embryos in vitro. Fertil Steril. 1996;66:394397. https://doi.org/10.1016/S0015–0282(16)58507-0Google Scholar
Barnes, FL, Crombie, A, Gardner, DK, et al. Blastocyst development and birth after in-vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching. Hum Reprod. 199;10:3243–3247. https://doi.org/10.1093/oxfordjournals.humrep.a135896Google Scholar
Biggers, JD, Racowsky, C. The development of fertilized human ova to the blastocyst stage in KSOM (AA) medium: is a two-step protocol necessary? Reprod Biomed Online. 2002;5:133140. https://doi.org/10.1016/S1472-6483(10)61615-XGoogle Scholar
Bontekoe, S, Johnson, N, Blake, D, Marjoribanks, J. Adherence compounds in embryo transfer media for assisted reproductive technologies: summary of a Cochrane review. Fertil Steril. 2015;103:14161417. https://doi.org/10.1016/j.fertnstert.2015.03.010Google Scholar
Borland, RM, Hazra, S, Biggers, JD, Lechene, CP. The elemental composition of the environments of the gametes and preimplantation embryo during the initiation of pregnancy. Biol Reprod. 1977;16:147157. https://doi.org/10.1095/biolreprod16.2.147Google Scholar
Brison, DR, Houghton, FD, Falconer, D, et al. Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover. Hum Reprod. 2004;19:23192324. https://doi.org/10.1093/humrep/deh409Google Scholar
Chatot, CL, Ziomek, CA, Bavister, BD, Lewis, JL, Torres, I. Random-bred 1-cell mouse embryos in vitro. J Reprod Fertil. 1989;86:679688.Google Scholar
Chronopoulou, E, Harper, JC. IVF culture media: past, present and future. Hum Reprod Update. 2015;21:3955. https://doi.org/10.1093/humupd/dmu040Google Scholar
Dieamant, F, Petersen, CG, Mauri, AL, et al. Single versus sequential culture medium: which is better at improving ongoing pregnancy rates? A systematic review and meta-analysis. JBRA Assist Reprod. 2017;21:240246. https://doi:10.5935/1518-0557.20170045Google Scholar
Dunglison, GF, Barlow, DH, Sargent, IL. Leukaemia inhibitory factor significantly enhances the blastocyst formation rates of human embryos cultured in serum-free medium. Hum Reprod. 1996;11:191196. https://doi.org/10.1093/oxfordjournals.humrep.a019016Google Scholar
Dyrlund, TF, Kirkegaard, K, Poulsen, ET, et al. Unconditioned commercial embryo culture media contain a large variety of non-declared proteins: a comprehensive proteomics analysis. Hum Reprod. 2014;29:24212430. https://doi.org/10.1093/humrep/deu220Google Scholar
Ebner, T, Shebl, O, Moser, M, Mayer, RB, Arzt, W, Tews, G. Group culture of human zygotes is superior to individual culture in terms of blastulation, implantation and life birth. Reprod Biomed Online. 2010;21:762768. https://doi.org/10.1016/j.rbmo.2010.06.038Google Scholar
Elder, K, Johnson, MH. The Oldham Notebooks: an analysis of the development of IVF 1969–1978. III. Variations in procedures. Reprod Biomed Soc Online. 2015;1:1933. https://doi.org/10.1016/j.rbms.2015.04.004Google Scholar
Fawzy, M, Emad, M, Elsuity, MA, et al. Cytokines hold promise for human embryo culture in vitro: results of a randomized clinical trial. Fertil Steril. 2019;112:849857.e1. https://doi.org/10.1016/j.fertnstert.2019.07.012Google Scholar
Fernández-Gonzalez, R, Ramirez, MA, Bilbao, A, De Fonseca, FR, Gutiérrez-Adán, A. Suboptimal in vitro culture conditions: an epigenetic origin of long-term health effects. Mol Reprod Dev. 2007;74:11491156. doi: 10.1002/mrd.20746. PMID: 17474101 DOI: 10.1002/mrd.20746Google Scholar
Fredrickson, J, Krisher, R, Morbeck, DE. The impact of the protein stabilizer octanoic acid on embryonic development and fetal growth in a murine model. J Assist Reprod Genet. 2015;32:15171524. https://doi.org/10.1007/s10815-015-0560-9Google Scholar
Gardner, DK. Mammalian embryo culture in the absence of serum or somatic cell support. Cell Biol Int. 1994;18:11631179. https://doi.org/10.1006/cbir.1994.1043Google Scholar
Gardner, DK. The impact of physiological oxygen during culture, and vitrification for cryopreservation, on the outcome of extended culture in human IVF. Reprod Biomed Online 2016;32:137141.Google Scholar
Gardner, DK, Lane, M. 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. Hum Reprod. 1996;11:27032712. https://doi.org/10.1093/oxfordjournals.humrep.a019195Google Scholar
Gardner, DK, Lane, M, Calderon, I, Leeton, J. Environment of the preimplantation human embryo in vivo: metabolite analysis of oviduct and uterine fluids and metabolism of cumulus cells. Fertil Steril. 1996;65:349353. https://doi.org/10.1016/s0015-0282(16)58097-2Google Scholar
Gardner, DK, Lane, M. Culture and selection of viable blastocysts: a feasible proposition for human IVF? Hum Reprod Update. 1997;3:367382. https://doi.org/10.1093/humupd/3.4.367Google Scholar
Gardner, DK, Lane, M. Culture of viable human blastocysts in defined sequential serum-free media. Hum Reprod. 1998;13 (Suppl 3), 148159. https://doi: 10.1093/humrep/13.suppl_3.148Google Scholar
Gardner, DK, Lane, MW, Lane, M. EDTA stimulates cleavage stage bovine embryo development in culture but inhibits blastocyst development and differentiation. Mol Reprod Dev. 2000;57:256261. https://doi.org/10.1002/1098-2795(200011)57:3<256::AID-MRD7>3.0.CO;2-PGoogle Scholar
Gardner, DK, Phil, D, Vella, P, Lane, M, Wagley, L, Schlenker, T. Culture and transfer of human blastocysts increases implantation rates and reduces the need for multiple embryo transfers. Fertil Steril. 1998;69:8488.CrossRefGoogle ScholarPubMed
Gómez, E, Carrocera, S, Martín, D, et al. Efficient one-step direct transfer to recipients of thawed bovine embryos cultured in vitro and frozen in chemically defined medium. Theriogenology. 2020;146:3947. https://doi.org/10.1016/J.THERIOGENOLOGY.2020.01.056Google Scholar
Hambiliki, F, Hanrieder, J, Bergquist, J, Hreinsson, J, Stavreus-Evers, A, Wånggren, K. Glycoprotein 130 promotes human blastocyst development in vitro. Fertil Steril. 2013;99:592599. https://doi.org/10.1016/j.fertnstert.2012.12.041Google Scholar
Hammond, J. Recovery and culture of tubal mouse ova. Nature. 1949;163:2829. https://doi.org/10.1038/163028b0Google Scholar
Hentemann, M, Mousavi, K, Bertheussen, K. Differential pH in embryo culture. Fertil Steril. 2011;95:12911294. https://doi.org/10.1016/j.fertnstert.2010.10.018Google Scholar
Iritani, A, Gomes, WR, Vandemark, NL. Secretion rates and chemical composition of oviduct and uterine fluids in ewes. Biol Reprod. 1969;1: 7276. https://doi.org/10.1095/biolreprod1.1.72Google Scholar
Iritani, A, Nishikawa, Y, Gomes, WR, VanDemark, NL. Secretion rates and chemical composition of oviduct and uterine fluids in rabbits. J Anim Sci. 1971;33:829835. https://doi.org/10.2527/jas1971.334829xCrossRefGoogle ScholarPubMed
Iritani, A, Sato, E, Nishikawa, Y. Secretion rates and chemical composition of oviduct and uterine fluids in sows. J Anim Sci. 1974;39:582588. https://doi.org/10.2527/jas1974.393582xGoogle Scholar
Kastrop, PMM, de Graaf-Miltenburg, LAM, Gutknecht, DR, Weima, SM. Microbial contamination of embryo cultures in an ART laboratory: sources and management. Hum Reprod. 2007;22:22432248. https://doi.org/10.1093/humrep/dem165Google Scholar
Kattal, N, Cohen, J, Barmat, LI. Role of coculture in human in vitro fertilization: a meta-analysis. Fertil Steril. 2008;90:10691076. https://doi.org/10.1016/j.fertnstert.2007.07.1349Google Scholar
Keith, B, Simon, MC. Hypoxia-inducible factors, stem cells, and cancer. Cell. 2007;129:465472. https://doi.org/10.1016/j.cell.2007.04.019Google Scholar
Kirkegaard, K, Hindkjaer, JJ, Ingerslev, HJ. Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring. Fertil Steril. 2013;99:738744.e4. https://doi.org/10.1016/j.fertnstert.2012.11.028Google Scholar
Kleijkers, SHM, Eijssen, LMT, Coonen, E, et al. Differences in gene expression profiles between human preimplantation embryos cultured in two different IVF culture media. Hum Reprod. 2015;30:23032311. https://doi.org/10.1093/humrep/dev179Google Scholar
Kleijkers, SHM, Mantikou, E, Slappendel, E, et al. Influence of embryo culture medium (G5 and HTF) on pregnancy and perinatal outcome after IVF: a multicenter RCT. Hum Reprod. 2016a;31:22192230. https://doi.org/10.1093/humrep/dew156Google Scholar
Kleijkers, SHM, Van Montfoort, APA, Bekers, O, et al. Ammonium accumulation in commercially available embryo culture media and protein supplements during storage at 2–8°C and during incubation at 37°C. Hum Reprod. 2016b;31:11921199. https://doi.org/10.1093/humrep/dew059Google Scholar
Koedooder, R, Mackens, S, Budding, A, et al. Identification and evaluation of the microbiome in the female and male reproductive tracts. Hum Reprod Update. 2019;25, 298325. https://doi.org/10.1093/humupd/dmy048Google Scholar
Lewis, WH, Gregory, PW. Cinematographs of living developing rabbit-eggs 1. Science. 1929;69:226229. https://doi.org/10.1126/science.69.1782.226-aGoogle Scholar
Lewis, N, Sturmey, RG. Embryo metabolism: what does it really mean? Anim Reprod. 2015;12, 521528.Google Scholar
Lippes, J, Enders, RG, Pragay, DA, Bartholomew, WR. The collection and analysis of human fallopian tubal fluid. Contraception. 1972;5:85103. https://doi.org/10.1016/0010-7824(72)90021-2Google Scholar
Lopata, A, Patullo, MJ, Chang, A, James, B. A method for collecting motile spermatozoa from human semen. Fertil Steril. 1976;27:677684. doi: 10.1016/s0015-0282(16)41899-6Google Scholar
Macklon, NS, Pieters, MHEC, Hassan, MA, Jeucken, PHM, Eijkemans, MJC, Fauser, BCJM. A prospective randomized comparison of sequential versus monoculture systems for in-vitro human blastocyst development. Hum Reprod. 2002;17:27002705.Google Scholar
Magli, MC, Gianaroli, L, Fiorentino, A, Ferraretti, AP, Fortini, D, Panzella, S. Fertilization and early embryology: improved cleavage rate of human embryos cultured in antibiotic-free medium. Hum Reprod. 1996;11:15201524. https://doi.org/10.1093/oxfordjournals.humrep.a019430Google Scholar
Mantikou, E, Bontekoe, S, van Wely, M, Seshadri, S, Repping, S, Mastenbroek, S. Low oxygen concentrations for embryo culture in assisted reproductive technologies. Hum Reprod Update 2012;19:209. doi: 10.1093/humupd/dms055.Google Scholar
Mantikou, E, Jonker, MJ, Wong, KM, et al. Factors affecting the gene expression of in vitro cultured human preimplantation embryos. Hum Reprod. 2016;31:298311. https://doi.org/10.1093/humrep/dev306Google Scholar
McLaren, A, Biggers, JD. Successful development and birth of mice cultivated in vitro as early embryos. Nature, 1958;182:877878. https://doi.org/10.1038/182877a0Google Scholar
Ménézo, Y. Synthetic medium for gamete survival and maturation and for culture of fertilized eggs[in French]. C R Acad Hebd Seances Acad Sci D. 1976;282:19671970.Google Scholar
Ménézo, Y, Lichtblau, I, Elder, K. New insights into human pre-implantation metabolism in vivo and in vitro. J Assist Reprod Genet. 2013;30:293303. https://doi.org/10.1007/s10815–013-9953-9Google Scholar
Ménézo, Y, Testart, J, Perrone, D. Serum is not necessary in human in vitro fertilization, early embryo culture, and transfer. Fertil Steril. 1984;42:750755. https://doi.org/10.1016/S0015–0282(16)48202-6Google Scholar
Moessner, J, Dodson, WC. The quality of human embryo growth is improved when embryos are cultured in groups rather than separately. Fertil Steril. 1995; 64:10341035. https://doi.org/10.1016/S0015–0282(16)57925-4Google Scholar
Morbeck, DE, Baumann, NA, Oglesbee, D. Composition of single-step media used for human embryo culture. Fertil Steril. 2017;107:10551060.e1. https://doi.org/10.1016/j.fertnstert.2017.01.007CrossRefGoogle ScholarPubMed
Morbeck, DE, Krisher, RL, Herrick, JR, Baumann, NA, Matern, D, Moyer, T. Composition of commercial media used for human embryo culture. Fertil Steril. 2014a;102:759766.e9. https://doi.org/10.1016/j.fertnstert.2014.05.043CrossRefGoogle ScholarPubMed
Morbeck, DE, Paczkowski, M, Fredrickson, JR, et al. Composition of protein supplements used for human embryo culture. J Assist Reprod Genet. 2014b;31:17031711. https://doi.org/10.1007/s10815–014-0349-2Google Scholar
Morgan, A, Babu, D, Reiz, B, Whittal, R, Suh, LYK, Siraki, AG. Caution for the routine use of phenol red – It is more than just a pH indicator. Chem Biol Interact. 2019;310:108739. https://doi.org/10.1016/j.cbi.2019.108739Google Scholar
Orsi, NM, Leese, HJ. Ammonium exposure and pyruvate affect the amino acid metabolism of bovine blastocysts in vitro. Reproduction. 2004;127:131140. https://doi.org/10.1530/rep.1.00031Google Scholar
Paternot, G, Debrock, S, D’Hooghe, TM, Spiessens, C. Early embryo development in a sequential versus single medium: a randomized study. Reprod Biol Endocrinol. 2010;8:83. https://doi.org/10.1186/1477-7827-8-83Google Scholar
Quinn, P, Kerin, JF, Warnes, GM. Improved pregnancy rate in human in vitro fertilization with the use of a medium based on the composition of human tubal fluid. Fertil Steril. 1985;44:493498. https://doi.org/10.1016/S0015–0282(16)48918-1Google Scholar
Restall, BJ. The fallopian tube of the sheep. I. Cannulation of the fallopian tube. Aust J Biol Sci. 1966;19:181186. https://doi.org/10.1071/bi9660181CrossRefGoogle ScholarPubMed
Restall, BJ, Wales, RG. The fallopian tube of the sheep. III. The chemical composition of the fluid from the fallopian tube. Aust J Biol Sci. 1966;19:687698.Google Scholar
Rodriguez-Wallberg, KA, Munding, B, Ziebe, S, Robertson, SA. GM-CSF does not rescue poor-quality embryos: secondary analysis of a randomized controlled trial. Arch Gyn Obstet. 2020;301:13411346. https://doi.org/10.1007/s00404–020-05532-3Google Scholar
Shao, WJ, Tao, LY, Xie, JY, Gao, C, Hu, JH, Zhao, RQ. Exposure of preimplantation embryos to insulin alters expression of imprinted genes. Comp Med. 2007;57:482486.Google Scholar
Sinclair, KD, Young, LE, Wilmut, I, McEvoy, TG. In-utero overgrowth in ruminants following embryo culture: lessons from mice and a warning to men. Hum Reprod. 2000;15(Suppl 5):68-86.Google Scholar
Sjöblom, C, Wikland, M, Robertson, SA. Granulocyte-macrophage colony-stimulating factor promotes human blastocyst development in vitro. Hum Reprod. 1999;14:30693076. https://doi.org/10.1093/humrep/14.12.3069Google Scholar
Steptoe, PC, Edwards, RG. Birth after the reimplantation of a human embryo. Lancet. 1978;312:366. https://doi.org/10.1016/S0140-6736(78)92957-4Google Scholar
Summers, MC, Bird, S, Mirzai, FM, Thornhill, A, Biggers, JD. Human preimplantation embryo development in vitro: a morphological assessment of sibling zygotes cultured in a single medium or in sequential media. Hum Fertil. 2013;16:278285. https://doi.org/10.3109/14647273.2013.806823Google Scholar
Sunde, A, Brison, D, Dumoulin, J, et al. Time to take human embryo culture seriously. Hum Reprod. 2016;31:21742780. https://doi.org/10.1093/humrep/dew157Google Scholar
Tarahomi, M, Vaz, FM, Straalen, J, et al. The composition of human preimplantation embryo culture media and their stability during storage and culture. Hum Reprod. 2019;34:14501461. https://doi.org/10.1093/humrep/dez102Google Scholar
Tay, JI, Rutherford, AJ, Killick, SR, Maguiness, SD, Partridge, RJ, Leese, HJ. Human tubal fluid: production, nutrient composition and response to adrenergic agents. Hum Reprod. 1997;12:24512456. https://doi.org/10.1093/humrep/12.11.2451Google Scholar
Tervit, HR, Whittingham, DG, Rowson, LE. Successful culture in vitro of sheep and cattle ova. J Reprod Fertil. 1972;30:493497. https://doi.org/10.1530/jrf.0.0300493Google Scholar
Van Montfoort, APA, Arts, EGJM, Wijnandts, L, et al. Reduced oxygen concentration during human IVF culture improves embryo utilization and cumulative pregnancy rates per cycle. Hum Reprod Open. 2020:hoz036. https://doi.org/10.1093/hropen/hoz036Google Scholar
Virant-Klun, I, Tomaževič, T, Vrtačnik-Bokal, E, Vogler, A, Krsnik, M, Meden-Vrtovec, H. Increased ammonium in culture medium reduces the development of human embryos to the blastocyst stage. Fertil Steril. 2006;85:526528. https://doi.org/10.1016/j.fertnstert.2005.10.018CrossRefGoogle Scholar
Wale, PL, Gardner, DK. The effects of chemical and physical factors on mammalian embryo culture and their importance for the practice of assisted human reproduction. Hum Reprod Update. 2016;22: 222. https://doi.org/10.1093/humupd/dmv034Google Scholar
Whitten, WK. Culture of tubal mouse ova. Nature. 1956;177:96. https://doi.org/10.1038/177096a0Google Scholar
Whittingham, DG. Fertilization of mouse eggs in vitro. Nature, 1968;220:592593. https://doi.org/10.1038/220592a0Google Scholar
Wydooghe, E, Vaele, L, Piepers, S, et al. Individual commitment to a group effect: strengths and weaknesses of Bovine embryo group culture. Reproduction. 2014;148:519529. https://doi.org/10.1530/REP-14-0213Google Scholar
Wydooghe, E, Vandaele, L, Heras, S, et al. Autocrine embryotropins revisited: how do embryos communicate with each other in vitro when cultured in groups? Biol Rev. 2017;92:505520. https://doi.org/10.1111/brv.12241Google Scholar
Young, LE, Sinclair, KD, Wilmut, I. Large offspring syndrome in cattle and sheep. Rev Reprod. 1998;3:155163. doi: 10.1530/ror.0.0030155. PMID: 9829550Google Scholar
Youssef, MMA, Mantikou, E, van Wely, M, et al. Culture media for human pre-implantation embryos in assisted reproductive technology cycles. Cochrane Database Syst Rev. 2015;20(11):CD007876.Google Scholar
Zhu, J, Li, M, Chen, L, Liu, P, Qiao, J. The protein source in embryo culture media influences birthweight: a comparative study between G1 v5 and G1-PLUS v5. Hum Reprod. 2014;29:13871392. https://doi.org/10.1093/humrep/deu103Google Scholar
Ziebe, S, Loft, A, Povlsen, BB, et al. A randomized clinical trial to evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium for in vitro fertilization. Fertil Steril. 2013;99:16001609.e2. https://doi.org/10.1016/j.fertnstert.2012.12.043Google Scholar

Save book to Kindle

To save this book to your Kindle, first ensure coreplatform@cambridge.org is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×