Embryo development of fresh versus vitrified metaphase II after intracytoplasmic sperm injection (ICSI): a sibling-oocyte study
| ISRCTN | ISRCTN60158641 |
|---|---|
| DOI | https://doi.org/10.1186/ISRCTN60158641 |
| Secondary identifying numbers | CVG02092008 |
- Submission date
- 18/05/2009
- Registration date
- 29/05/2009
- Last edited
- 21/12/2009
- Recruitment status
- No longer recruiting
- Overall study status
- Completed
- Condition category
- Pregnancy and Childbirth
Prospectively registered
Protocol
Statistical analysis plan
Results
Individual participant data
Plain English summary of protocol
Not provided at time of registration
Contact information
Dr Laura Rienzi
Scientific
Scientific
G.EN.E.R.A.
Clinica Valle Giulia
Via de Notaris 2B
Rome
00197
Italy
| rienzi@generaroma.it |
Study information
| Study design | Randomised active controlled parallel group trial |
|---|---|
| Primary study design | Interventional |
| Secondary study design | Randomised controlled trial |
| Study setting(s) | Hospital |
| Study type | Treatment |
| Participant information sheet | Not available in web format, please use the contact details below to request a patient information sheet |
| 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 |
| Study objectives | 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(s) | Local medical ethics committee (Clinica Valle Giulia, Roma) approved on the 1st September 2008 |
| Health condition(s) or problem(s) studied | 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 |
| Primary outcome measure | 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 study start date | 02/09/2008 |
| Completion date | 10/03/2009 |
Eligibility
| Participant type(s) | Patient |
|---|---|
| Age group | Adult |
| Sex | Female |
| Target number of participants | 40 |
| Key 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. |
| Key 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 |
| Date of first enrolment | 02/09/2008 |
| Date of final enrolment | 10/03/2009 |
Locations
Countries of recruitment
- Italy
Study participating centre
G.EN.E.R.A.
Rome
00197
Italy
00197
Italy
Sponsor information
G.EN.E.R.A. - Clinica Valle Giulia (Italy)
Hospital/treatment centre
Hospital/treatment centre
via de Notaris 2B
Rome
00197
Italy
| rienzi@generaroma.it | |
| Website | http://www.generaroma.it/ |
"ROR" |
https://ror.org/05aq4y378 |
Funders
Funder type
Hospital/treatment centre
G.EN.E.R.A. - Clinica Valle Giulia (Italy)
No information available
Results and Publications
| Intention to publish date | |
|---|---|
| Individual participant data (IPD) Intention to share | No |
| IPD sharing plan summary | Not provided at time of registration |
| Publication and dissemination plan | Not provided at time of registration |
| IPD sharing plan |
Study outputs
| Output type | Details | Date created | Date added | Peer reviewed? | Patient-facing? |
|---|---|---|---|---|---|
| Results article | results | 01/01/2010 | Yes | No |
