Embryo culture medium which is the best

The protein supplement and concentration used in the culture media should also be accounted for. Protein supplements can be extremely variable and impact embryo development. In an ideal scenario, the laboratory would supplement the same protein to each medium tested to control for protein or lot number variations.

Comparisons of culture media should also ensure the same type of dish is used between the two media examined. Ideally, when comparing media, both treatments should be placed within the same incubator. As mentioned previously, this requires careful assessment of pH to ensure the gas conditions are appropriate for both media tested. If the same CO 2 concentration is not possible for both media, then separate incubators must be used. In this case, careful attention to other variables must be taken i.

Once the media comparison is underway, an interim assessment is likely warranted. Following completion of the trial, careful examination of the data should be done to determine if laboratory or clinical outcomes justify a medium change or not. The criteria for justifying a change should have been determined prior to the onset of the trial.

Jason E. Allow Cookies. Join Our Mailing List Today! Comparing Embryo Culture Media. August 13, Trialing a new embryo culture medium is a common occurrence in the IVF laboratory.

Define endpoints Prior to starting the media comparison, the endpoints need to be clearly defined and powered for the difference anticipated. Use a sibling oocyte split study design The recommended method to compare culture media is a sibling oocyte split.

Validate your setup Key considerations when comparing media include first validating the culture system to ensure the medium is being used within the manufacturer's recommended parameters. Addition of LIF to embryo culture media has been shown to enhance the quality of human embryos Dunglison et al.

Glycoprotein Gp is involved in mediating the effect of a range of cytokines and the addition of Gp to embryo culture media might have a positive effect embryo development in vitro Hambiliki et al. Animal studies have shown that the presence of granulocyte-macrophage colony-stimulating factor GM-CSF in culture media may exert a positive effect on embryo development, leading to significant interest in this molecule as a putative supplement to embryo culture.

However, in a large prospective randomized study, the addition of GM-CSF to the embryo culture media revealed no overall positive effect on embryo development, implantation rates, or delivery rates. However, a post hoc subgroup analysis indicates a possible positive effect in the subgroup of patients with previous failed cycles Ziebe et al.

However, whether suppression of apoptosis, which represents a mechanism by which an embryo may eliminate damaged cells, is of benefit is unclear. A more recent prospective randomized study compared the outcome after culturing human embryos either in a standard medium or the same medium with addition of a mixture of cytokines LIF, GM-CSF, heparin-binding epidermal growth factor HB-EGF.

The addition of cytokines to the medium improved embryo quality Fawzy et al. A few manufacturers add insulin to some of their culture media intended for IVF.

The presence of insulin has been demonstrated to alter the profile of DNA methylation in mice embryos Shao et al. DNA methylation is one way of regulating expression of genes and a change in DNA-methylation pattern is therefore usually accompanied with a change m-RNA profile and the proteins synthesized.

Whether such effects present in human embryos remains unknown. Thus, the effectiveness of supplementation of culture media with growth factors and cytokines remains unclear. In vivo, the genital tract contains gonadal steroids, such as estradiol and progesterone.

These steroids are key regulators of genital tract function and influence the molecules that are secreted into the fallopian tube lumen and to the uterine cavity. Addition of steroid hormones to embryo culture media has not proven to be beneficial. A major challenge of steroid supplementation is the difficulty of dissolving them in culture media, a feature shared with other lipophilic substances.

In vivo, they are predominantly bound to carriers, such as serum albumin and sex hormone binding globulin SHBG. Some embryo culture media are very complex and reflective of products intended for continuous culture of mammalian cells. Such formulations can include up to 80 different components, including fatty acids, vitamins, iron chelators, trace metals, and intermediates in metabolism.

It has not been shown that these very complex culture media are superior to less complex media and it still unknown to what extent these varied components will affect development of human embryos in vitro. Hyaluronic acid may be involved in implantation and adding it to the medium used to transfer the embryo to the uterus has been shown to have a moderate positive effect on implantation rates Bontekoe et al.

Most embryo culture media contain bicarbonate and pH is regulated by the CO 2 concentration in the incubator. The CO 2 concentration should be adjusted to give a pH in the physiological range 7. One study on mouse embryos indicates a beneficial effect of having a higher pH at the zygote stage and lower pH from the cleavage stage Hentemann, Addition of a pH indicator, such as phenol red, may offer a visual check on the pH of the culture media.

However, phenol red is not an inert molecule and may change the metabolism of cultured mammalian cells Morgan et al. It is therefore perhaps advisable to use phenol red—free culture media. Follicular aspirates often contain traces of vaginal flora and it is reasonable to assume that most semen samples are not sterile Kastrop et al. Antibiotics are, therefore, added to culture media, even though it has been demonstrated that presence of antibiotics may reduce embryo quality Magli et al.

These antibiotics are usually not manufactured with assisted reproduction in mind, and it can be a challenge for media manufacturers to source antibiotics that do not contain trace amounts of molecules, such as lipopolysaccharides LPS , that will have negative effect on embryo development. It is a matter of discussion whether to add penicillin in combination with streptomycin or gentamicin or to use gentamicin alone.

Semen and vaginal flora contain both Gram-positive and Gram-negative bacteria and since penicillin primarily works against Gram-positive bacteria, it is necessary to add streptomycin or gentamicin to also inhibit the growth of Gram-negative bacteria.

Vaginal flora may include Candida species. Embryo culture media does not contain antimycotics, and if vaginal fluid is aspirated during oocyte recovery, the cultures may be heavily contaminated with Candida cells. Some culture media contain EDTA which will bind divalent cations.

In some animal models, the presence of EDTA will be beneficial for embryo development. For human embryo culture using modern culture media, it is not established whether the presence of EDTA is necessary for optimal embryo development.

Many embryo culture media contain surface tension and viscosity modulators to enable simpler handling of small volumes of media.

The effect, if any, on the development of human embryos is unknown. Chapter 5 — Culture Media and Embryo Culture. Theriogenology , 71 7 , 20 Jan Cited by: 25 articles PMID: Hum Reprod , 24 4 , 14 Jan Cited by: 23 articles PMID: Fertil Steril , 91 5 , 25 Apr Cited by: 44 articles PMID: Jansen RP. Intern Med J , 35 2 , 01 Feb Cited by: 9 articles PMID: Gynecol Obstet Fertil , 34 9 , 28 Aug Cited by: 2 articles PMID: Contact us. Europe PMC requires Javascript to function effectively.

Recent Activity. Search life-sciences literature Over 39 million articles, preprints and more Search Quick link: Coronavirus articles and preprints. Recent history Saved searches. Abstract Available from publisher site using DOI. A subscription may be required. Michelle Lane Search articles by 'Michelle Lane'. Lane M ,. Gardner DK. Affiliations All authors 1. Share this article Share with email Share with twitter Share with linkedin Share with facebook. Abstract With the growing move in in-vitro fertilization IVF clinics to transfer fewer embryos to women, there is an increasing reliance on the IVF laboratory to maximize embryo viability.

Subsequently, there is justified scrutiny on the culture system and the media used to sustain the human embryo in vitro. The transfer of fewer embryos to patients also creates an increased dependence on the ability to cryopreserve embryos successfully. Therefore, in addition to the ability of a culture system to produce a single top-quality embryo for transfer, it is also necessary to enhance the cryotolerance of sibling embryos so that they can survive freezing or vitrification.

Therefore, when examining which culture media is the best, it is prudent to not only examine the ability of a culture system to produce a pregnancy with the one or two highest-grade embryos, but also to determine how many embryos from the entire cohort both fresh and frozen embryos are capable of producing a live birth. Additionally, research on animal models has demonstrated that stress, and the resultant adaptation to conditions during pre-implantation stages, can affect pregnancy loss and fetal growth.

It is therefore important to understand the role of each medium component and to identify possible sources of cellular stress to the embryo that will ultimately affect the function and viability of the conceptus. Full text links Read article at publisher's site DOI : References Articles referenced by this article Physiology and culture of the human blastocyst.