GAIN medium

Single-step culture medium

GAIN medium is a bicarbonate buffered medium optimized to provide an excellent in vitro environment for human gametes and embryos. GAIN medium can be used for sperm preparation, oocyte incubation and fertilization. It is a single-step medium designed for continuous embryo culture from day 1 to expanded blastocyst stage. The medium can also be used for embryo transfer. Oocyte and embryo culture should be performed in a CO₂ incubator.

The medium contains 0.35% human serum albumin (EMA, Ph Eur, USP, FDA approved), gentamicin and low concentrations of phenol red (pH indicator). Information about the composition of the product can be found in the material safety data sheet.

Regulatory

Europe: CE-marked – Brazil: registered

Product order codes

GAIN010 : GAIN medium – 1x 10mL
GAIN020 : GAIN medium – 5x 20mL

Product 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
Shelf life	12 months from date of produce
Albumin	FDA (USA) and EMA (Europe) compliant

Key components

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 about 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, et al., Protein Supplementation of Human IVF Culture Media., Journal of Assisted Reproduction and Genetics (2002),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)

Frequently Asked Questions Certificate Of Analysis

Oocyte Sperm Processing Fertilization Embryo Transfer