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
Dont 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.
Tratamiento farmacológico. PsicofármacosPILAR ALEJANDRA SÁIZ MARTÍNEZ(1); Mª PAZ GARCÍA-PORTILLA GONZÁLEZ(1); SARA MARTÍNEZ BARRONDO(2); Mª TERESA BASCARÁN FERNÁNDEZ(2, 3); BEGOÑA PAREDES (1) Profesor Titular de Psicología Médica. (2) Colaborador de Honor – Dpto. de Medicina - Área de Psiquiatría. (3) Doctorando Área de Psiquiatría. Lugar de Trabajo: Área de Psiquiatría