Blastocoele FluidNCT01427413
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
An observational study of Blastocoele Fluid, run by Cervesi Hospital, Cattolica, Italy at 1 site in Italy.
Trial summary
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. Read more about this study
- Who is it for?
- Ages 18 to 45 years, women only
- Conditions the study is for
- Blastocoele Fluid
- How many people
- 100 participants
- Where
- 1 siteItaly
- Study dates
- Sep 2011to Dec 2023
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 45 years
- Women only
- No healthy volunteers
- Adult
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.
Who may be able to join
- Ovarian hyperstimulation disease
- Blastocyst formation
Who cannot join
None listed.
Trial locations and contacts
1 site in 1 country.
Italy1 site, 1 recruiting
- Recruiting
Cervesi Hospital
Cattolica, Rimini, 47841
Silvia De Stefani, Biotecnology, +39 320 1111937, silvia.destefani83@libero.it
Simone Palini, biology, +39 339 4572101, simonepalini@yahoo.it
About this study
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.
- Conditions
- Blastocoele Fluid
- Keywords
- blastocoele fluid, mass spectrometry, metabolomics
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
| Study group | Role | What they receive |
|---|---|---|
| Hyper1 | Only patients who achieved hyperstimulation pathology after external administration of gonadotrophin during IVF treatment |
What is being measured
The primary outcome is the main question the study is designed to answer.
standardize the method of aspiration
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.
Measured: 2 months
metabolite detection
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.
Measured: 6 months
Secondary outcomes (1)
analysis of the correlation between blastocyst morphology and metabolic profile in the blastocoele fluid
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
Measured: 1 year
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted1 Sep 2011 (estimated)
- Study startSep 2011
- Primary completionJan 2022 (estimated)
- Study completionDec 2023 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Blastocoele FluidStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Case only
- Time perspective
- Retrospective
- Enrolment
- 100 participants (estimated)
Sponsor and investigators
- Lead sponsor
- Cervesi Hospital, Cattolica, Italy
- Sponsor type
- University, hospital or other
- Collaborators
- Università degli Studi La Tuscia, Tecnobios Riproduzione, Istituto Clinico Humanitas
- Responsible party
- Principal investigator, Palini Simone, Cervesi Hospital, Cattolica, Italy
- Investigators
- Simone Palini, biology, Study director, Cervesi Hospital, Cattolica, Italy
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT01427413
- Sponsor's protocol ID
- SS4e
- Status last verified
- Jul 2020
- First submitted
- 29 Aug 2011
- Last updated
- 9 Feb 2021
- Results posted
- No
- Data monitoring committee
- Yes
Study at a glance
From the registry record, last updated by the sponsor on 9 Feb 2021. Recruitment status is as the sponsor reported it: confirm with the study team.
- 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
- ClinicalTrials.gov ID
- NCT01427413
- Acronym / sponsor ID
- SS4e
- Conditions
- Blastocoele Fluid
- Keywords
- blastocoele fluid, mass spectrometry, metabolomics
- Study type
- Observational
- Recruitment status
- Unknown status
- Enrolment
- 100 participants (estimated)
- Lead sponsor
- Cervesi Hospital, Cattolica, ItalyUniversity, hospital or other
- Collaborators
- Università degli Studi La Tuscia, Tecnobios Riproduzione, Istituto Clinico Humanitas
- Study start
- Sep 2011
- Primary completion
- Jan 2022 (estimated)
- Study completion
- Dec 2023 (estimated)
- First posted
- 1 Sep 2011 (estimated)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
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.
Study design
- Observational model
- Case only
- Time perspective
- Retrospective
- Number of arms
- 1
| Arm | Type | Receives |
|---|---|---|
| Hyper1 | – | –Only patients who achieved hyperstimulation pathology after external administration of gonadotrophin during IVF treatment |
Who may qualify
- Ages
- 18 to 45 years
- Sex
- Female only
- Healthy volunteers
- Not accepted
- Age groups
- Adult
- 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.
Key inclusion criteria (2 listed)
- Ovarian hyperstimulation disease
- Blastocyst formation
Key exclusion criteria (0 listed)
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Primary outcome
standardize the method of aspiration
Time frame: 2 months
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
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.
1 secondary outcome
analysis of the correlation between blastocyst morphology and metabolic profile in the blastocoele fluid
Time frame: 1 year
Results and safety
Results are not available in the source record.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 9 Feb 2021.
Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Simone Palini, biology study director
Cervesi Hospital, Cattolica, Italy
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Blastocoele Fluid, run by Cervesi Hospital, Cattolica, Italy at 1 site in Italy.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 2 inclusion and 0 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: status not confirmed, as of the sponsor's last update on 9 Feb 2021. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Italy. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
Results are not available in the source record.
How can a doctor refer a patient or contact the research team?
Contact Simone Palini, biology (+39 339 4572101, simonepalini@yahoo.it), or a site near the patient. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
standardize the method of aspiration (time frame: 2 months). The study lists 2 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 9 Feb 2021.