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GAIN™ medium

2 step fertilisation and IVF medium
Fertilization EmbryoHandling EmbryoTransfer

GAIN medium consists of 2 types of ready to use cell culture media for use with human embryos and gametes. The media can be used for fertilization, zygotes and embryos culture through the cleavage stages and up to the expanded blastocyst stage and for embryo transfer.

  • GAIN medium Early Stage (GAIN ES) can be used for the handling of oocytes (in preparation of, or during IVF/ICSI) and for cell culture from fertilization up to 4-cell stage, for the processing of semen and for embryo transfer.

  • GAIN medium Blastocyst (GAIN BL) is designed for embryo culture from 4-cell up to expanded blastocyst stage and for embryo transfer. Alternatively, the medium can be used immediately after fertilization, up till expanded blastocyst stage.


  • The media contain 0.35% human serum albumin (EMA, Ph Eur, USP, FDA approved), gentamicin and low concentrations of phenol red (pH indicator).

    GAIN media undergo strict quality control to comply with the following specifications:

    pH 37°C - 5% CO2 7.20-7.45
    Osmolality 270-290mOsm/kg
    Sterility Sterile
    Endotoxine < 0.25 EU/mL
    Mouse embryo test ≥ 80% blastocysts after 96h incubation
    Shelflife 12 months from date of produce
    Albumin FDA (USA) and EMEA (Europe) compliant

    GAIN ES and GAIN BL are CE marked as Class III medical devices according to EU directive 93/42/EEC.


    Product order codes

    GAIN010ES 1x 10ml GAIN medium Early Stage
    GAIN010BL 1x 10ml GAIN medium Blastocyst


    The product composition can be found in the MSDS (see Resources). Additional information on some components is provided below:

    Component Benefit
    No HEPES (0 mM)

    GAIN medium does not contain HEPES, therefore CO2 incubation is required.

    Gentamicin sulphate (10 mg/L)

    GAIN medium is also available with the addition of gentamicin sulphate.

    Indeed, even under aseptic conditions, commensal bacteria are easily introduced in culture media. Since these contaminations can be detrimental to gametes, embryos and ART outcome, a large number of customers choose to use media supplemented with gentamicin (Quinn 2014).

    Minimal Inhibitory Concentration (MIC) tests done by FertiPro N.V. have demonstrated that embryo culture media supplemented with 0.010 g/L gentamicin efficiently eliminate bacterial growth.

    Even though, each handling should be performed under strict aseptic conditions (LAF-bench)!

    EDTA

    EDTA probably acts in several ways (Quinn 2000). EDTA is known to act as a chelator of heavy metal divalent cations that may be present as trace contaminants in media components and plastic culture ware.
    Moreover, EDTA acts through the chelation of iron resulting in a lowered generation of reactive oxygen species (Nasr-Esfahani et al. 1990). EDTA may also suppress glycolysis and may therefore be beneficial for the culture of cleavage stage embryos (Lane and Gardner 2001).


    HSA (3.5 g/L)

    GAIN medium contains 3.5 g/Liter Human Serum Albumin (HSA) to optimize medium performances. HSA is universally added to most ART media because it is widely considered to be of benefit.

    The role of albumin in ART media is extensive, including:
    - Stabilization of the cell membrane of the embryo in the medium (Malda, et al. 2008).
    - Inhibition of lipid peroxidation that can be damaging to sperm (Alvarez en Storey 1995).
    - Carrier and source of essential molecules needed by the embryo (Malda, et al. 2008).
    - Detoxification by binding waste products from cell metabolism (Blake, et al. 2004)
    - Facilitating gamete/embryo manipulation by preventing adsorption to the surface through saturation of potential binding sites (Blake, et al. 2004)


    Literature concerning the components


         Alvarez, JG., and BT. Storey, Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa., Mol Reprod Dev. (1995),Vol.42,No.3,pp.334-46
         Blake, D., P. Svalander, M. Jin, C. Silversand, and L. Hamberger., Protein Supplementation of Human IVF Culture Media., Journal of Assisted Reproduction and Genetics (2004),Vol.19,No.3,pp.137-143
         Lane, Gardner, Inhibiting 3-phosphogluycerate kinase by EDTA stimulates the development for the cleavage stage mouse embryo., Mol. Reprod dev (2001),Vol.60,No.2,pp.233-240
         Malda, J. et al., Cell Nutrition, In Tissue engineering, by C. Van Blitterswijk. London, UK: Academia Press, Elsevier, (2008),pp.327-362
         Nasr-Esfahani, M., M.H. Johnson, and R.J. Aitken, The effect of iron and iron chelators on the in-vitro block to development of the mouse preimplantation embryo: BAT6 a new medium for improved culture of mouse embryos in vitro., Human Reproduction (1990),Vol.5,No.8,pp.997-1003
         Quinn, Review of Media Used in ART Laboratories., Journal of Andrology (2000),Vol.21,No.5,pp.610-615
         Quinn, Media and embryo interactions - anitbiotics, In Culture media, Solutions, and Systems in Human ART, by P. Quinn, 14. United Kingdom: Cambridge University Press (2014)

    Resources

    Click on the links below for more information.