Condition category
Urological and Genital Diseases
Date applied
16/01/2012
Date assigned
01/03/2012
Last edited
14/08/2014
Prospective/Retrospective
Prospectively registered
Overall trial status
Completed
Recruitment status
No longer recruiting

Plain English Summary

Lay summary under review 2

Trial website

Contact information

Type

Scientific

Primary contact

Dr Stephan Gordts

ORCID ID

Contact details

Leuven Institute for Fertility and Embryology (LIFE)
Tiensevest 168
Leuven
3000
Belgium

Additional identifiers

EudraCT number

ClinicalTrials.gov number

Protocol/serial number

N/A

Study information

Scientific title

Evaluation of blastocyst survival following vitrification and thawing using two methods - Fast Freeze™ Kit based on S3 and the Vit. Kit®: a randomised controlled study

Acronym

Study hypothesis

To evaluate and compare the survival rate of vitrified-warmed blastocysts submitted to the Global® Fast Freeze Kit – based on S3 (LifeGlobal, Guelph, Canada) and the Vit. Kit® protocol (Irvine Scientific, Santa Ana, CA). Furthermore, a slight modification of the S3 rapid freezing protocol was tested. We expect to observe differences in terms of morphology, kinetics and cell survival on the three groups of blastocysts submitted to the different cryopreservation protocols.

Ethics approval

Not provided at time of registration

Study design

Single-centre randomised controlled study

Primary study design

Interventional

Secondary study design

Randomised controlled trial

Trial setting

Hospitals

Trial type

Screening

Patient information sheet

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

Condition

Reproductive medicine

Intervention

Group 1: Global® Fast Freeze and Thawing media – based on S31 blastocyst at the time is removed from the incubator and transferred with a micropipette to previously made drops of vitrification solutions. The blastocyst is then transferred to a drop of solution 3 and immediately loaded in a 0.25 or 0.30 ml previously labelled freezing straw with solution S3, attached to a straw holding and aspiration device. The blastocyst is kept in a column of S3 solution, between two air bubbles. The straw is then sealed with an appropriate heat sealer and held vertically together with a thermocouple probe, to be lowered into the mouth of the liquid nitrogen tank. Once the temperature reaches -100 degree C, the straw is held still for 2 (0.25 cc straws) or 3 minutes (0.30 cc straws), before it is plunged into liquid nitrogen and stored in the cryotank.

Group 2
Some of the blastocysts are cryopreserved with the method described above, but with slight modifications. Once sealed, the straw is plunged directly into liquid nitrogen, without the "intermediate step of holding the straw for 2 or 3 minutes at -100 degree C. The remaining steps remain unchanged and thawing is performed similarly to the procedure decrcribed above.

Gropu 3: Vit Kit® media- (Irvine Scientific)
One blastocyst at the time is removed from the incubator and transferred to a equilibration solution drop, where it should stay for 5-15 minutes, until it shrinks and returns to its size. Then the blastocyst is transferred consecutively to 4 drops of vitrification solutions, at intervals of 5 seconds. Using a cryotip attached to a 1ml syringe, the blastocyst is aspirated. The cryotip is sealed in both ends and the cover sleeve is slid back to protect the tip. The covered cryotip is then plunged directly into liquid nitrogen. Thawing is done in a water bath at 37ºC for 3 seconds and the content of the cryotip is then expelled and mixed with 1ul of thawing solution for 1 minute. Subsequently the blastocyst is placed in another drop of thawing solution for 1 minute, then on 2 drops of dilution solution and finally the blastocyst is washed 3 times. The blastocyst is then transferred to pre-equilibrated culture dish containing drops of Global® media.

Group 4 (control group: non vitrified day 5 and day 6 blastocysts)

Intervention type

Other

Phase

Not Applicable

Drug names

Primary outcome measures

1. Survival assessment by vital staining
2. Assessment of cell number and survival rate following vitrification
3. Thawing and culture overnight is performed with the Life-dye TM and propidium iodide (Life-dead cell staining kit; Biovision, CA, US). Briefly blastocysts are incubated in a buffer media with 1ul of each staining solutions for 15 min in the dark, at 37ºC.

Secondary outcome measures

1. Differences in terms of morphology, kinetics and cell survival on the three groups. Digital images of each blastocyst (day 5 or day 6) are acquired before the vitrification procedure, immediately after thawing and after overnight culture, using a digital still camera (Octax, Microscience Gmb H, Bruckberg, Germany) mounted on an inverted optical microscope (TE2000-S, Nikon Eclipse, Japan), with a thermal control microscope stage (MS100, Linkam, Surray, UK).
2. Assessment of blastocyst morphology quality is done according to the classification developed by Gardner & Schoolcraft 1999 and re-expansion is assessed following overnight culture.

Overall trial start date

01/03/2012

Overall trial end date

01/02/2013

Reason abandoned

Eligibility

Participant inclusion criteria

1. Blastocysts derived from 1PN and 3PN embryos (abnormal embryos)
2. Blastocysts derived from biopsied and rejected embryo
3. Blastocysts derived from too fast or too slow embryos (that were neither transferred nor cryopreserved)
4. Blastocysts derived from culture of embryos (days 2 and 3) donated for scientific research

Participant type

Patient

Age group

Adult

Gender

Female

Target number of participants

100

Participant exclusion criteria

Still useful embryos and normal embryos not donated for scientific research

Recruitment start date

01/03/2012

Recruitment end date

01/02/2013

Locations

Countries of recruitment

Belgium

Trial participating centre

Leuven Institute for Fertility and Embryology (LIFE)
Leuven
3000
Belgium

Sponsor information

Organisation

Leuven Institute for Fertility and Embryology (LIFE) (Belgium)

Sponsor details

Tiensevest 168
Leuven
3000
Belgium

Sponsor type

Industry

Website

Funders

Funder type

Industry

Funder name

Leuven Institute for Fertility and Embryology (LIFE) (Belgium)

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

Publication citations

Additional files

Editorial Notes