The outcome of an assisted reproduction treatment
(ART) cycle is highly dependent on numerous
factors including the clinic location, equipment
and disposables used (
These products are commercially available, their
quality is meticulously tested, and they are readyto-
use. Therefore the use of these high quality
products should significantly improve embryo
quality, cost-efficiency of the treatment, and the
clinic’s reputation (
The purpose of this study was to compare the efficiency of two different commercially available media products (ISM1, MediCult, Denmark vs. G-1TM v5, Vitrolife, Sweden) in supporting the development of embryos (days 2 and 3), their ability to implant and deliver a healthy baby at term.
This prospective randomized study was performed
during a five month period (February-
June, 2009). During this period all culture
conditions, routines and disposables were kept
constant in the laboratory. On the day of oocyte
pick up, there were 583 patients randomized
to both groups according to a randomization
list based on sequential numbers in sealed envelopes.
The ISM1 group (experimental) consisted
of 293 patients compared to 290 patients
in the routine lab procedure group (control).
Depending on the causes of infertility, patients
received either standard
Inclusion criteria were counseling patients undergoing ICSI treatment, between 18 to 40 years of age who had a minimum of 2 follicles (≥18 mm) at the time of oocyte collection.
Patients were randomly assigned to have their fertilized oocytes cultured in ISM1 (MediCult, cycles: n=293, written informed consent was taken from all the couples who received this culture medium) or routine lab procedure (G-1TM v5; vitrolife, cycles: n=290) according to the daily medium schedule for oocyte retrieval. This study was approved by Ethical Committee of Royan Institute.
Down regulation and ovarian stimulation were
performed in accordance with a previously reported
stimulation regimen (
Once ovarian suppression was confirmed (serum E2 ≤50 pg, FSH ≤12 IU and LH ≤5 IU), ovarian stimulation was initiated with recombinant FSH (Gonal F; SC injection, 150 IU/day, Serono, Switzerland). The dose was adjusted according to the response of the ovarian follicular development and monitored by serial vaginal ultrasonography. When at least three follicles reached 18 mm in diameter, both GnRH agonist and hMG were discontinued and a single dose of hCG (10000 IU; Pregnyl; Organon, Netherlands) was administered.
Oocyte retrieval was performed via vaginal ultrasound- guided follicle aspiration, 36-38 hours after administration of hCG.
After egg collection, the cumulus-oocyte complexes were either placed in 50 μL droplets
of Universal IVF medium and covered with
liquid paraffin (MediCult, Denmark) as the
experimental group or in 50 μL droplets of
Ham’s F-10, (Biochrom AG; Berlin, Germany),
which had been supplemented with 10%
recombinant human serum albumin (rHA, Vitrolife)
and covered with pre-incubated mineral
oil (Sigma, St. Louis, MO), as the control
group. Oocyte denudation was performed
with ICSI Cumulase (MediCult) in the experimental
group or with 80 IU of hyaluronidase,
(Sigma, USA) in the control group, two hours
after egg collection. Sperms were immobilized
in 10% PVP droplets (MediCult) in the experimental
group and in10% PVP droplets (Global)
in the control group by breaking the tail by
an injection pipette. Then, a single sperm with
head front was injected into each oocyte that
had a visible first polar body (MII) (
The fertilization rate was controlled 18-20 hours after sperm injection. Normally fertilized oocytes that had evidence of two pronuclei were washed and transferred to overnight pre-incubated in 20 μl droplets of ISM1 or G- 1TM v5 supplemented with 10% recombinant human serum albumin in the experimental and control groups respectivly and covered with either pre-incubated liquid paraffin (experimental group) or mineral oil (control group). All cultures incubated at 37˚C, in 5% O2 and 6% CO2.
On the day of embryo transfer (44-72 hours after injection), we scored embryo morphology according to the following quality criteria:
Day 2: 2-4 even size blastomeres with ≤10% fragmentation
Day 3: 6-8 even size blastomeres with ≤10% fragmentation
Day 2: 2-4 even or uneven size blastomeres with 10%-20% fragmentation
Day 3: 6-8 even or uneven size blastomeres with 10%-20% fragmentation
Uneven and few blastomeres with >20% fragmentation
Based upon the embryo quality, female age and number of previous cycles, we selected a maximum of four high quality embryos and incubated them in UTM (MediCult) for the experimental group or Embryo Glue (Vitrolife) for the control group, for a period of 20-120 minutes before embryo transfer.
All embryo transfers were performed with a Labotect catheter (Labotect, Germany) against a filled urine bladder with the expertise of gynecologists and embryologists.
We used the Statistical Package for the Social Sciences (SPSS 11.5; Chicago, IL, USA, http:// www.spss.com) software to analyze differences amongst the variables of both groups by Chisquare and Fisher’s exact tests for categorical variables or the student’s t test for continuous variables, as appropriate. A p value <0.05 was considered to be statistically significant.
Patient characteristics of the two groups are summarized in table 1. The number of cycles, mean female age, indications for infertility, number of previous treatment cycles, retrieved oocytes and percentage of cleaved embryos were similar in both groups.
There were no significant differences between
the groups in terms of cleavage rates, number
of embryos transferred, clinical pregnancy and
implantation rates and multiple pregnancies.
There were a significantly higher percentage
of excellent embryos in the experimental group
(42.7%; ISM1) compared to the control group
(39%, p<0.05). There was also a trend towards
higher clinical and multiple pregnancy rates as
well as implantation rates (
In terms of abortion, the experimental group (20.5%) was not significantly different from the control group (21.1%). There was no significant difference in baby take home rates (BTHR) between both groups, however both the length [48.8 cm (experimental) vs. 46.0 cm (control)] and weight [3.03 kg (experimental) vs. 2.66 kg (control)] of babies born were significantly higher in the expreimental group (p<0.001 for both parameters).
Patients’ ART cycle characteristics in the MediCult (experimental) and lab routine media (control)
|Parameters||Experimental group||Control group||P value|
|32.4 ± 5.7||31.8 ± 5.6||0.18|
|170/293 (58%)||170/290 (58.6%)||0.88|
|66/293 (22.5%)||62/290 (21.4%)||0.73|
|44/293 (15%)||46/290 (16%)||0.77|
|13/293 (4.4%)||12/290 (4.1%)||0.83|
|1.7 ± 1.2||1.8 ± 1.0||0.21|
|9.1 ± 5.6||8.8 ± 5.1||0.52|
Fertilization, cleaved embryo, embryo quality and clinical pregnancy in the MediCult (experimental) and lab routine media (control) groups
|Parameters||Experimental group||Control group||*P value|
|86% (1471/1706)||88% (1455/1678)||0.68|
|43% (628)||39% (567)||0.04a|
|39% (580)||41% (599)||0.33|
|18% (261)||20% (288)||0.15|
|2.4 ± 0.8||2.4 ± 0.8||0.83|
|32.1% (94/293)||27.6% (80/290)||0.23|
|15% (102/697)||12% (84/693)||0.16|
|8.5% (8/94)||3.8% (3/80)||0.19|
|91.5% (86/94)||96.2% (77/80)||0.19|
|8.5% (8/94)||2.5% (2/80)||0.09|
|0% (0/94)||1.3% (1/80)||0.46|
a; Statistically significant differences between the two groups and *; Fisher’s exact test.
Clinical outcome, abortion and take-home baby rate in the MediCult (experimental) and lab routine media (control) groups
|Parameters||Experimental group||Control group||*P value|
|20.5% (18/88)||21.1% (16/76)||0.9|
|77.8% (14/18)||87.5% (14/16)||0.58|
|16.7% (3/18)||12.5% (2/16)||0.48|
|5.5% (1/18)||0% (0/16)||0.35|
|84.3% (86/102)||80.4% (78/97)||0.57|
|8.5% (8/94)||3.8% (3/80)||0.19|
|78.9% (56/71)||66.7% (40/60)||0.1|
|21.1% (15/71)||31.7% (19/60)||0.1|
|0% (0/71)||1.7% (1/60)||0.1|
|44.1% (38/86)||51.3% (40/78)||0.3|
|55.8% (48/86)||48.7% (38/78)||0.3|
|48.8 ± 0.52||46.0 ± 0.66||0.001 a|
|3.03 ± 0.07||2.66 ± 0.08||0.001 a|
a; Statistically significant differences between the two groups and *; t test.
Of all pregnant patients, 10 (n=6 for expreimental group and n=4 for control group) patients were not available and were excluded from table 3.
There are a variety of different
A general agreement does not exist regarding
the efficacy of different commercially available
Several studies have investigated the effect of
ISM1 and G-1TM v5 media on embryo quality and
implantation rate. Embryo morphology on days 2
and 3 significantly enhanced when the embryos
were cultured in GIII series versus ISM1 (
A prospective study on sibling oocytes showed
that embryo quality in the day 2 stage improved
under ISM1 culture rather than FertiCult culture
medium, even if data regarding pregnancy and
implantation rate were not reported (
For this reason, we conducted a prospective randomized trial at Royan Institute to compare embryo development parameters, IVF outcome and BTHR at the end of the gestational period between ISM1 and G-1TM v5 media, which is used routinely in our laboratory.
The result of the study suggests that both ISM1
and G-1TM v5 support embryo development
Good embryo quality, cleavage rate, and single, triplet pregnancy rate in the control group were higher than ISM1 medium, but there were no significant differences. In ISM1, the implantation, clinical pregnancy and twin pregnancy rates were higher than the G-1TM v5 medium. However, this difference was not significant. The percentage of excellent embryos was significantly higher than the control group which might be reflected in the characteristics of babies born. Babies born after culture in ISM1 had both significantly higher birth weight and were also longer compared to the control group, both of which were significant.
In vivo as well as
Although in vivo physiologic reactive oxygen
species production is controlled continuously by
a strong cellular defense, the lack of this system
Singleton children born after IVF have a significantly
lower birth weight compared with their
spontaneously conceived peers (
"The possible interference of assisted reproduction
techniques (ART) with epigenetic reprogramming
during early embryo development has recently
sparked renewed interest about the reported
lower birth weight among infants born as a consequence
of infertility treatments" (
Both the shorter duration of pregnancy and the
reduced neonatal birth weight could be caused by
inactivation of paternal alleles during the imprinting
process. If the shorter duration of pregnancy
and the lower birth weight after IVF and ICSI can
be attributed to a distinct defect of epigenetic phenomena
at work during parental imprinting, then
this defect may have later effects on the individual
ISM1 culture medium was formulated to promote
correct imprinting for ensuring correct development
and implantation (
This study suggests that on the one hand ISM1 contains antioxidant elements and on the other hand it is formulated to promote correct imprinting to ensure correct development and implantation. ISM1 is presumed to be a more effective culture medium that generates higher quality embryos, which is reflected in the characteristics of babies born. Since numerous factors that include mother’s diabetes and multiple pregnancies, among others may also impact the height and the weight of the newborn, we cannot directly relate the change in these two characteristics solely to the culture media. For reaching a definitive conclusion, additional studies that consider these factors are warranted.