Condition category
Pregnancy and Childbirth
Date applied
18/05/2009
Date assigned
29/05/2009
Last edited
21/12/2009
Prospective/Retrospective
Retrospectively registered
Overall trial status
Completed
Recruitment status
No longer recruiting

Plain English Summary

Not provided at time of registration

Trial website

Contact information

Type

Scientific

Primary contact

Dr Laura Rienzi

ORCID ID

Contact details

G.EN.E.R.A.
Clinica Valle Giulia
Via de Notaris 2B
Rome
00197
Italy
rienzi@generaroma.it

Additional identifiers

EudraCT number

ClinicalTrials.gov number

Protocol/serial number

CVG02092008

Study information

Scientific title

Embryo development of fresh versus vitrified metaphase II after intracytoplasmic sperm injection (ICSI): a prospective randomised active-controlled parallel group sibling-oocyte study

Acronym

Study hypothesis

Non-inferiority trial in order to evaluate the effectiveness of the oocyte vitrification procedure, effectiveness being defined by fertilisation rate after intracytoplasmic sperm injection (ICSI) per warmed oocyte.

Ethics approval

Local medical ethics committee (Clinica Valle Giulia, Roma) approved on the 1st September 2008

Study design

Randomised active controlled parallel group trial

Primary study design

Interventional

Secondary study design

Randomised controlled trial

Trial setting

Hospitals

Trial type

Treatment

Patient information sheet

Not available in web format, please use the contact details below to request a patient information sheet

Condition

Intracytoplasmic sperm injection cases, oocyte vitrification

Intervention

After oocyte denudation, MII oocytes with normal morphology were randomly allocated to fresh ICSI insemination or to vitrification procedure. If pregnancy was not obtained a subsequent ICSI cycle was performed with warmed oocytes of the same cohort. In both groups, 3 oocytes were inseminated per cycle by ICSI procedure.

The vitrification and warming procedures were performed according to Kuwayama and co-Authors (2005). Commercial kits were used (Vitrification and Warming KIT, Kitazato BioPharma Co, Japan).

The vitrification procedure was performed at room temperature (RT). Oocytes were equilibrated in the equilibration solution (ES) containing 7.5% ethylene glycol (EG) and 7.5% dimethylsulfoxide (DMSO) in HEPES buffered basic culture medium M-199 with 20% synthetic serum substitute (SSS). To perform the equilibration gradually, the oocytes were first placed in a 20 microlitre drop of M199+20%SSS and, immediately, after mixed with a second 20 microlitre drop of ES. After 3 minutes incubation, a third 20 microlitre drop of ES solution was mixed. Finally, the oocyte were moved in a pure drop of 20 microlitre ES and incubated for an additional 6 - 9 minutes. The oocytes (1 to 3, contemporaneously) were then transferred in 1 ml of vitrification solution (VS) containing 15% EG, 15% DMSO and 0.5M sucrose in M199+20%SSS for 1 minute. The oocytes were then placed on the Cryotop strip in a single small drop of VS. Much care was driven to re-aspirate, as much as possible, the excess of VS in such way to leave just a thin layer around each oocyte. The Cryotop was then immediately submerged into liquid nitrogen. Finally, the plastic cap was pulled over the Cryotop inside the liquid nitrogen and the sample was stored submerged in liquid nitrogen.

The first step of warming procedure was performed at 37°C. The cap was removed in liquid nitrogen and the cryotop was immediately submerged in 1 ml of warming solution containing 1.0M sucrose in M199+20%SSS. After 1 minute, oocytes were placed in 1 ml solution containing 0.5M sucrose, and incubated at RT for 3 minutes. Finally, the oocytes were washed at RT for 6 minutes in 2 different dishes containing 1 ml basic medium M199+20%SSS each, and transferred into 1 ml culture media. Degenerated oocytes were removed from the cohort.

The surviving oocytes were co-cultured at 37°C (6% CO2 and 5%O2) for exactly 2 hours before ICSI.

Intervention type

Other

Phase

Not Applicable

Drug names

Primary outcome measures

Non-inferiority in fertilisation rates calculated per warmed and per injected oocyte, assessed 16 - 18 hours post-treatment (ICSI).

Secondary outcome measures

1. Pronuclear morphology
2. Embryo development, assessed 42 - 44 hours post-treatment
3. Patient's baseline characteristics
4. Clinical outcomes

Overall trial start date

02/09/2008

Overall trial end date

10/03/2009

Reason abandoned

Eligibility

Participant inclusion criteria

Between September 2008 and February 2009 consecutive patients not older than 42 years of age, presenting greater than 6 normal appearing metaphase II (MII) oocytes and undergoing ICSI treatment with ejaculated sperm in the Centre for Reproductive Medicine GENERA in Rome.

Participant type

Patient

Age group

Adult

Gender

Female

Target number of participants

40

Participant exclusion criteria

1. Female partner older than 42 years old
2. Less than 6 normal appearing MII oocytes retrieved
3. Surgically extracted spermatozoa
4. Very severe oligoastenoteratozoospermia (motile sperm count less than 500,000/ml after preparation)
5. Patients enrolled in our polar body biopsy programme

Recruitment start date

02/09/2008

Recruitment end date

10/03/2009

Locations

Countries of recruitment

Italy

Trial participating centre

G.EN.E.R.A.
Rome
00197
Italy

Sponsor information

Organisation

G.EN.E.R.A. - Clinica Valle Giulia (Italy)

Sponsor details

via de Notaris 2B
Rome
00197
Italy
rienzi@generaroma.it

Sponsor type

Hospital/treatment centre

Website

http://www.generaroma.it/

Funders

Funder type

Hospital/treatment centre

Funder name

G.EN.E.R.A. - Clinica Valle Giulia (Italy)

Alternative name(s)

Funding Body Type

Funding Body Subtype

Location

Results and Publications

Publication and dissemination plan

Not provided at time of registration

Intention to publish date

Participant level data

Not provided at time of registration

Results - basic reporting

Publication summary

2010 results in http://www.ncbi.nlm.nih.gov/pubmed/19861328

Publication citations

  1. Results

    Rienzi L, Romano S, Albricci L, Maggiulli R, Capalbo A, Baroni E, Colamaria S, Sapienza F, Ubaldi F, Embryo development of fresh 'versus' vitrified metaphase II oocytes after ICSI: a prospective randomized sibling-oocyte study., Hum. Reprod., 2010, 25, 1, 66-73, doi: 10.1093/humrep/dep346.

Additional files

Editorial Notes