Embryo culture
Directions for supplementation of un-supplemented G-SeriesTM media
Day 0
G-1™ PLUS
OVOIL™
Prepare micro-droplet culture
dishes with 25 µL droplets of
G-1 PLUS for washing and for
culture. Cover with OVOIL and
pre-equilibrate at
37°C 6 % CO2
overnight
G-MOPS™ PLUS
Warm G-MOPS PLUS (for fer-
tilisation assessment) in rinsed
tightly capped tubes in a warm-
ing incubator without CO2* at
37°C overnight
* An adequately calibrated
warming block can be used
for tubes instead of a warming
incubator.
Ensure that the denudation
and washing procedures
are performed at 37°C
1.160906.6
Day 1 If your clinic is located at a
higher altitude than sea level,
1. Fertilisation assessment
CO2 percentage should be
For inseminated oocytes,
increased, see graph below.
transfer the oocytes to a centre
12.0 %
well dish with pre-warmed 11.0 %
G-MOPS PLUS. If denudation 10.0 %
and fertilisation assessment can 9.0 %
be performed within 2 minutes, 8.0 %
G-IVF™ PLUS can be used 7.0 %
instead of G-MOPS PLUS.
6.0 %
Remove cumulus and corona
5.0 %
0m 1000 m 2000 m
cells from oocytes using a
denudation pipette and Day 2
assess fertilisation at Assessment
37°C Assess embryo cleavage.
For embryo transfer day 2,
see separate Embryo transfer
For ICSI oocytes, assess fertili- protocol
sation in the G-1 PLUS micro-
droplet culture dish.
Day 3
Assessment
2. Culture
Assess embryo cleavage.
Wash the zygotes
For embryo transfer day 3,
extensively in
see separate Embryo transfer
the G-1 PLUS
protocol
micro-droplet
dishes prepared
on Day 0 and transfer the
zygotes to 25 µL G-1 PLUS We recommend
culture droplets covered with G-MOPS PLUS for
OVOIL. Culture at washing and assess-
ment of embryos before
37°C 6 % CO2 transfer to G-2 PLUS
overnight or for 2 days or EmbryoGlue®
can be found in the G-Series Manual on www.vitrolife.com.
Once supplemented, the media should be used as the
G-Series PLUS media described below.
Blastocyst culture Day 4
1. Prepare micro-well dishes Prepare micro-droplet
for Blastocyst culture culture dishes
G-2™ PLUS G-2 PLUS
OVOIL OVOIL
In the morning of Prepare centre
day 3, prepare well dishes with
3000 m 4000 m 5000 m micro droplet fresh G-2 PLUS.
culture dishes with Prepare micro-droplet
25 µL droplets of dishes for prolonged culture if
G-2 PLUS for washing and for needed and pre-equilibrate at
culture. Cover with OVOIL and
pre-equilibrate at 37°C 6 % CO2
overnight
37°C 6 % CO2 ≥ 6 h
Day 5
In the morning of day 5
2. Move embryos to
Assess embryo
G-2™ PLUS
cleavage, and
In the afternoon move the blastocysts
of day 3, wash
selected for transfer
the embryos
and cryo preservation to the
extensively in
equilibrated G-2 PLUS
equilibrated G-2 PLUS centre well dishes and leave at
droplets and transfer the
embryos to G-2 PLUS culture 37°C 6 % CO2
droplets, maximum 5 embryos until 10-30 min
per droplet. Culture at before transfer
37°C 6 % CO2
2 days For blastocyst transfer, see
separate Embryo transfer
protocol