Viewing Study NCT06084377



Ignite Creation Date: 2024-05-06 @ 7:38 PM
Last Modification Date: 2024-10-26 @ 3:11 PM
Study NCT ID: NCT06084377
Status: COMPLETED
Last Update Posted: 2023-10-18
First Post: 2023-10-11

Brief Title: Isolation of Cell Free Fetal DNA From Spent Culture Medium and Its Potential Role for Preimplantation Genetic Diagnosis PGD
Sponsor: Damascus University
Organization: Damascus University

Study Overview

Official Title: Isolation of Extracellular Embryonic DNA FromIn Vitro Fertilization IVF Embryonic Culture Medium and Its Potential Role for Preimplantation Genetic Screening PGS
Status: COMPLETED
Status Verified Date: 2023-10
Last Known Status: None
Delayed Posting: No
If Stopped, Why?: Not Stopped
Has Expanded Access: False
If Expanded Access, NCT#: N/A
Has Expanded Access, NCT# Status: N/A
Acronym: None
Brief Summary: Preimplantation genetic testing for chromosomal screening is at present the most reliable method to assess the genetic status of preimplantation embryos DNA isolated and amplified from trophectoderm TE biopsiesis the method currently used for preimplantation genetic testing but it has significant limitations There is increasing evidence for a true noninvasive approach consisting of the analysis of cell-free DNA cfDNA released by the embryo into the spent blastocyst medium SBM during the late stages of preimplantation development

This study is to assess whether Noninvasive prenatal genetic diagnosis using cell-free fetal DNA in spent culture medium will substitute for trophectoderm biopsy for heritable disorders screening and if it represents the total fetal genomic DNA

A total of 27 spent blastocyst media SBM from women undergoing IVF will be enrolled with agesless than44 years whose indications for preimplantation genetic diagnosis for EuploidAneuploid embryo determination were advanced maternal age recurrent miscarriage or recurrent implantation failureWe examined the presence of the cell free DNA in spent culture media and whether this DNA is reliable and can be representative of the chromosomal constitution of a blastocyst So we can affirm that Noninvasive prenatal determination of fetal sex using cell-free fetal DNA used here as a marker provides an alternative to invasive techniques for some heritable disorders
Detailed Description: Noninvasiveness is a common theme across medical specialties in the 21st century In obstetrics noninvasive prenatal testing provoked a paradigm shift in prenatal clinical practice through the sequencing of fetal cell-free DNA cfDNA in maternal plasma However current practice for preimplantation genetic testing for aneuploidy PGT-A requires trophectoderm biopsy which entails technical challenges and embryo viability concerns The possibility to overcome these issues reached the horizon in recent years with the discovery that cfDNA released by the embryo to the culture media during the latest stages of development offers a new strategy to assess the preimplantation embryo genetically

Our study evaluates the possibility to isolate fetal cell-free DNA cfDNA from culture media and compare the results with TE biopsy samples from the same patients who underwent an IVF stimulation cycle with intracytoplasmic sperm injection at Al-Sharq Hospital between March 2022 and April 2023 During the studys time frame all embryos were cultured until the blastocyst stage All enrolled patients underwent niPGD for gender determination using spent culture media as well as standard PGD with TE biopsy using Polymerase chain reaction technique for SRY gene The embryos were individually cultured in 30-mL media droplets and embryo development progressed using sequential culture media consistent with routine laboratory practice For niPGD analysis the blastocyst-stage culture media on Day 5 was collected 10-20 µl immediately before TE biopsy was performed Only embryos achieving a grade 1 fragmentation less than 15 were considered eligible for biopsy and vitrification The embryo culture media samples underwent whole-genome amplification WGA using the DOPlify WGA kit PerkinElmer to generate representative amplification of total DNA from spent culture media The DOP-PCR-based WGA reproducibly amplifies total DNA to produce microgram quantities of amplified and purified genomic DNA in less than 3 hours We aim to evaluate the concordance rates between trophectoderm biopsy results and cfDNA collected from spent media with an optimized protocol to obtain an adequate quantity and quality of genomic DNA This interesting work is an important effort to establish the feasibility of the new noninvasive approach that can be a reliable alternative for chromosomal abnormalities assessment of in vitro cultured embryos which will lead to the birth of a living and healthy baby

Later on we will perform the SRY gene detection for sex determination using the Polymerase Chain Reaction on the cell-free fetal DNA isolated from spent media samples and compare our results with the TB results

Study Oversight

Has Oversight DMC: None
Is a FDA Regulated Drug?: False
Is a FDA Regulated Device?: False
Is an Unapproved Device?: None
Is a PPSD?: None
Is a US Export?: None
Is an FDA AA801 Violation?: None