Directions for use for ferticult ivf medium

In open systems such as with the Falcon or Nunc dishes, about 1 mL of medium is placed in each well.
CELL CULTURE MEDIUM FOR IN VITRO CULTURE OF HUMAN EMBRYOS, A further 3 mL is placed in the reservoir surrounding the wells.
This helps to maintain humidity as well as providing medium with which to wash the eggs and embryos.
FertiCult IVF medium is sterilized by aseptic filling STERILE A Up to five eggs per dish (or well if using Nunc dishes) may be cultured together, usually with about 100.000 sperm per well.
So if each patient during the next day is expecting 15 eggs or so at egg collection, at least six dishes per patient should be prepared (three for initial holding and cleaning of eggs during collection and then three dishes for overnight culture).
FertiCult IVF medium is a ready to use formulation for the in When using Nunc dishes, two would be enough (since there are vitro culture of mammalian embryos. It is designed for short 4 wells per dish, the resulting 8 wells would be more than term culture only (up to 48 hours in culture).
The medium is complete and needs no further additives.
If preferred 10 % patient serum v/v can be added.
The dishes are then placed in the incubator to equilibrate FertiCult IVF medium is suitable for microdroplets under oil or overnight at 37°C and under an atmosphere of 5% C0 in air.
equally suited to culture in Falcon 3037 organ culture dishes As with the microdroplets method described above, equilibra- tion overnight is highly recommended.
As with all IVF media, FertiCult IVF medium ought to be pre- In the open system, paraffin oil is not necessary, but sometimes incubated in the incubator for 24 hours before use (with lid a 1mL layer of mineral oil may be added over the medium in the inner well. When using such an open system the incubator must After equilibration overnight, the prepared dishes are ready for use. During egg collection, eggs are identified in the follicular aspirates and then washed before placing into the wells or droplets in the dish (as appropriate). Once egg collection is over, eggs will be checked and may be rewashed, before then being placed into fresh droplets/wells for overnight culture.
Generally, within 6 hours of egg collection, eggs will be insemi- nated with prepared sperm and then left overnight in the incuba- The procedures described above are for the first 24 hours of cul- ture. Once embryos have been identified by the presence of two pronuclei, they are usually placed into fresh dishes containing fresh In the past, where people have used cord serum or patient serum, the initial medium contains 10% and the changeover (after fertili- PRODUCT SPECIFICATIONS AND QUALITY CONTROL When using FertiCult IVF medium no extra protein is required at FertiCult IVF medium is manufactured according to these specifica- changeover. Dishes should be prepared as described above, equilibrated overnight, after which they will be ready to receive the NB. Test tube culture is now rare, but for those still using this method, place 1mL of FertiCult IVF medium in each tube and then follow the same procedure as for Falcon or Nunc dishes.
- Mouse-embryo test: > 70% blastocysts after 96 hours in Store FertiCult IVF medium between 2-25°C.
- Use of Ph Eur or USP grade products if applicable Keep from light.
WARNING AND PRECAUTION All human, organic material should be considered potentially For microdroplets, between 100-250µL of FertiCult IVF medium Handle all specimens as if capable of transmitting HIV or may be dispensed around the culture dish, up to 8 per 60mm dish.
hepatitis. Always wear protective clothing when handling speci- The dish is then filled with 5 mL of pre-washed and pre-equilibrated light mineral oil (nontoxic and preferably embryo tested, e.g.
FertiCult IVF medium does not contain antibiotics, always work under strict hygienic conditions (laminar flow) to avoid contami- One egg is usually placed with about 10.000 sperm per micro- nation, or add your own antibiotics (penicillin) at about 100 units droplet, so if each patient during the next day is expecting 16 eggs, at least two dishes per patient should be prepared. The Don’t use FertiCult IVF medium if cloudy.
dishes are then placed in the incubator (usually non-humidified) to equilibrate overnight at 37°C and under an atmosphere of 5% This overnight equilibration is highly recommended.

Source: http://www.cryocatalog.ru/freezer/FertiCult_IVF.pdf