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FertiCult™ Flushing medium

HEPES buffered cell culture medium for use in IVF
SpermProcessing IUI OocyteHandling EmbryoHandling EmbryoTransfer

FertiCult Flushing medium is a formulation for washing of human ova, spermatozoa and embryos. FertiCult Flushing medium can also be used for swim-up techniques of human spermatozoa, sperm injection in oocytes during ICSI, introduction of washed spermatozoa in the uterus (IUI) and for embryo transfer.

All the products in the FertiCult™ product range undergo strict quality control to comply with the following specifications:

 pH 7.3-7.6
 Osmolality 270-290mOsm/kg
 Sterility Sterile
 Endotoxin < 0.25EU/ml
 Mouse embryo assay ≥ 80% blastocysts after 96h incubation
 Shelf life 18 months from date of produce
 Albumin FDA (USA) and EMEA (Europe) compliant

FertiCult™ Flushing medium is CE marked as a Class 3 medical device according to the European Medical Device Directive.


Product order codes

FLUSH020 FertiCult Flushing medium - 5x 20ml
FLUSH050 FertiCult Flushing medium - 5x 50ml
FLUSH100 FertiCult Flushing medium - 3x 100ml
FLUSH020PHR FertiCult Flushing medium w/ ph.red - 5x 20ml
FLUSH050PHR FertiCult Flushing medium w/ ph.red - 5x 50ml
FLUSH100PHR FertiCult Flushing medium w/ ph.red - 3x 100ml
FLUSH020PRG FertiCult Flushing medium w/ ph.red & gentamicin - 5x 20ml
FLUSH050PRG FertiCult Flushing medium w/ ph.red & gentamicin - 5x 50ml
FLUSH100PHR_G FertiCult Flushing medium w/ ph.red & gentamicin - 3x 100ml


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

Component Benefit
HEPES (18 mM)

HEPES stabilizes the pH under air (Clark and Swain 2014), therefore CO2 incubation is not required.

HSA (4g/L)

FertiCult Flushing medium contains 4 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)

Phenol red sodium salt (0.003 g/L)

FertiCult Flusing medium is also available with the addition of phenol red.

Phenol red functions as a visual pH indicator:
- > 7.35: the medium is colored red to purple.
- 7.15 - 7.35: the medium is colored pink to rose.
- < 7.15: the medium is colored yellow.

0.003 g/L is the minimum concentration of phenol red needed to view a color (Fleming and Cooke 2008). pH stabilization is essential when handling gametes or embryos. Therefore, validation of pH changes is strongly recommended. Changes in medium color also act as an indicator for bacterial or fungal contamination. Infection affects pH and this can easily be observed in medium containing a pH indicator such as phenol red.

Gentamicin sulphate (0.010 g/L)

FertiCult Flushing 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)!


Product literature


     Bandularatne E., Bongso A., Evaluation of Human Sperm Function After Repeated Freezing and Thawing, Journal of Andrology (2002),Vol.23,No.2,pp.242-249
     Marchetti, C.; Obert, G.; Deffosez, A.; Formstecher, P.; Marchetti, P., Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm., Hum. Reprod. (2002),Vol.17,No.5,pp.1257-65
     Parmegiani, L.; Cognigni, GE.; Bernardi, S.; Troilo, E.; Taraborelli, S.; Arrone, A.; Maccarini, AM.; Filicori, M., Comparison of two ready-to-use systems designed for sperm-hyaluronic acid binding selection before intracytoplasmic sperm injection: PICSI vs. Sperm Slow: a prospective, randomized trial., Fertil. Steril. (2012),Vol.98,No.3,pp.632-7
     Perrin, J.; Saias-Magnan, J.; Lanteaume, A.; Serment, G.; Grillo, J-M.; Karsenty, G., Amélioration de la qualité du sperme vésical en présence d'éjaculation rétrograde: apport de l'instillation vésicale pré-éjaculatoire de FertiCult., Abstract book congres FFER (2009),Vol.0
     Pongsuthirak, P.; Vutyavanich, T., Developmental competence of human embryos derived from in vitro maturation of immature oocytes retrieved during cesarean section., J. Obstet Gynaecol. Res (2014),Vol.40,No.2,pp.459-64
     Pont, JC.; Patrat, C.; Fauque, P.; Camp, ML.; Gayet, V.; Wolf, JP., Pre-washing catheter dramatically improves the post intrauterine insemination pregnancy rate., Gynecol. Obstet. Fertil. (2012),Vol.40,No.6,pp.356-9
     Tongue,E.; Var, T.; Onalan, G.; Altinbas, S.; Tokmak, A.; Karakas, N.; Gulerman, C., Comparison of the effectiveness of single versus double intrauterine insemination with three different timing regimens., Fertil. Steril. (2010),Vol.94,No.4,pp.1267-70


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
     Clark, N.A. and Swain, J.E., Buffering systems in IVF., Culture media, Solutions, and Systems in Human ART, by P. Quinn (2014),pp.30-46
     Fleming, S., Cooke, S., 4.3.7 Use of coloured pH-indicators - advantages and disadvantages., In Textbook of Assisted Reproduction for Scientists in Reproductive Technology; Australia: Vivid Publishing (2008),pp.78-79
     Malda, J. et al., Cell Nutrition, In Tissue engineering, by C. Van Blitterswijk. London, UK: Academia Press, Elsevier, (2008),pp.327-362
     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.