Now Available for Public Comment: Notice of Proposed Rulemaking (NPRM) for FDAAA 801 and NIH Draft Reporting Policy for NIH-Funded Trials

Identification and Standardization of a Method That Would Allow the Study of the Metabolic Profile of Blastocoele Lays the Foundation to Assess Blastocyst Metabolomic Profile and Its Relation With Embryo Morphology and Embryo Implantation

This study is currently recruiting participants. (see Contacts and Locations)
Verified August 2011 by Cervesi Hospital, Cattolica, Italy
Sponsor:
Collaborators:
Università degli Studi La Tuscia
Tecnobios Riproduzione
Istituto Clinico Humanitas
Information provided by (Responsible Party):
Palini Simone, Cervesi Hospital, Cattolica, Italy
ClinicalTrials.gov Identifier:
NCT01427413
First received: August 29, 2011
Last updated: July 30, 2013
Last verified: August 2011
  Purpose

While the number of assisted reproduction cycles increases worldwide, the introduction of actual technological improvements in the ability to quickly and non-invasively identify the best embryos for transfer still represents a critical goal for reproductive medicine. Indeed, embryo assessment is currently performed through the analysis of morphology and cleavage rate. Recent studies have sought to identify a correlation between qualitative-quantitative profiles of small molecules of metabolic interest and the outcome of embryo transfer. Some of these molecules seem to be best suited for this purpose, including glucose, lactate, pyruvate or amino acid levels. Approaches relying on both optical and non-optical spectroscopy have been proposed to non-invasively monitor the embryo culture media. However, the non-invasive approach only offers an indirect strategy to monitor embryos and a turn-around solution to bypass the limits of detection of these analytical techniques. In this paper the investigators pave the way for direct assessment of embryos through the mass spectrometry-based analysis of blastocoele fluid, which is withdrawn from the blastocoele cavity prior to cryostorage of blastocysts. The investigators show how it is possible to detect most of the already documented metabolites of interest right at the very heart of the blastocyst, without disrupting the workflow of a classic laboratory pipeline.


Condition
Blastocoele Fluid

Study Type: Observational
Study Design: Observational Model: Case-Only
Time Perspective: Retrospective
Official Title: Identification and Standardization of a Method That Would Allow the Study of the Metabolic Profile of Blastocoele Lays the Foundation to Assess Blastocyst Metabolomic Profile and Its Relation With Embryo Morphology and Embryo Implantation

Further study details as provided by Cervesi Hospital, Cattolica, Italy:

Primary Outcome Measures:
  • standardize the method of aspiration [ Time Frame: 2 months ] [ Designated as safety issue: Yes ]

    The method for blastocyst micropuncturing and aspiration of blastocoel fluid is according also to the last literature about blastocyst vitrification. In brief, expanded day 5 blastocysts were removed from culture and transferred to a 30 nl droplet of pre-warmed Hepes buffer.

    An injection pipette was introduced avoiding contaminations through the trophectoderm, and blastocoel fluid was aspirated until the blastocyst had fully collapsed around the pipette. The retrieved fluids were expelled into new purified water drops and frozen at −80°C alongside 0.5 nl control droplets of purified water.


  • metabolite detection [ Time Frame: 6 months ] [ Designated as safety issue: Yes ]
    metabolite detection through rapid resolution reversed phase (RR-RP) high performance liquid chromatography (HPLC)-mass spectrometry (MS). From sample volumes as low as 0.5 nl we could detect and quantify against external standards a group of metabolites, whose roles in blastocyst development and embryo metabolism have long been postulated. The list included i) ATP adenosine triphosphate ; ii) glucose-6-phosphate; iii) lactate; iv) NAD+ nicotinamide-adenine dinucleotide+ and v) NADH and vi) NADPH nicotinamide-adenine dinucleotide phosphate; vii) 6-phosphogluconic acid; viii) glutamic acid and ix) α-ketoglutarate.


Secondary Outcome Measures:
  • analysis of the correlation between blastocyst morphology and metabolic profile in the blastocoele fluid [ Time Frame: 1 year ] [ Designated as safety issue: Yes ]
    blastocyst classify according to morphological criteria in the book "Atlas of Human Blastocyst" by L. Veeck and associate, to each morphological class a characteristic metabolic profile, in order to scientifically validate the observational and objective criteria up to now used in our lab


Biospecimen Retention:   Samples With DNA

blastocoele fluid


Estimated Enrollment: 100
Study Start Date: September 2011
Estimated Study Completion Date: December 2015
Estimated Primary Completion Date: January 2014 (Final data collection date for primary outcome measure)
Groups/Cohorts
Hyper1
Only patients who achieved hyperstimulation pathology after external administration of gonadotrophin during IVF treatment

  Eligibility

Ages Eligible for Study:   18 Years to 45 Years
Genders Eligible for Study:   Female
Accepts Healthy Volunteers:   No
Sampling Method:   Probability Sample
Study Population

All the patients that are unable to be subjected to embryo transfer due to her ovarian hyperstimulation. In those cases the embryos will be vitrified and during this procedure we collapse the blastocyst sucking the fluid.

Criteria

Inclusion Criteria:

  • Ovarian hyperstimulation disease
  • Blastocyst formation
  Contacts and Locations
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the Contacts provided below. For general information, see Learn About Clinical Studies.

Please refer to this study by its ClinicalTrials.gov identifier: NCT01427413

Contacts
Contact: Simone Palini, biology +39 339 4572101 simonepalini@yahoo.it
Contact: Silvia De Stefani, Biotecnology +39 320 1111937 silvia.destefani83@libero.it

Locations
Italy
Cervesi Hospital Recruiting
Cattolica, Rimini, Italy, 47841
Contact: Silvia De Stefani, Biotecnology    +39 320 1111937    silvia.destefani83@libero.it   
Contact: Simone Palini, biology    +39 339 4572101    simonepalini@yahoo.it   
Sponsors and Collaborators
Cervesi Hospital, Cattolica, Italy
Università degli Studi La Tuscia
Tecnobios Riproduzione
Istituto Clinico Humanitas
Investigators
Study Director: Simone Palini, biology Cervesi Hospital, Cattolica, Italy
  More Information

No publications provided

Responsible Party: Palini Simone, Embryologist, Cervesi Hospital, Cattolica, Italy
ClinicalTrials.gov Identifier: NCT01427413     History of Changes
Other Study ID Numbers: SS4e
Study First Received: August 29, 2011
Last Updated: July 30, 2013
Health Authority: Italy: Comitato Etico di Area Vasta Romagna e I.R.S.T.

Keywords provided by Cervesi Hospital, Cattolica, Italy:
blastocoele fluid
mass spectrometry
metabolomics

ClinicalTrials.gov processed this record on November 20, 2014