A Prospective Study to Evaluate the Gender Prediction of Blastocysts by Using Cell-free DNA Within a Culture Medium

Overview

Journal Int J Reprod Biomed

Specialty Reproductive Medicine

Date 2022 Oct 3

PMID 36187736

Authors

Somayeh Barzanouni

Farideh Moramezi

Mahvash Zargar

Hamid Galehdari

Masoud Hemadi

Affiliations

Soon will be listed here.

Abstract

Background: Preimplantation genetic diagnosis (PGD) has been used as an option for couples with the possibility of having a baby with a genetic disorder. The common method for performing this test involves isolating 1 cell from day 3 or a few cells from day 5 embryos and performing genetic studies on the cell-extracted DNA. This method is invasive and can cause abortion after implantation in the uterus. Because of this, 2 noninvasive methods for performing a PGD have been studied: PGD using blastocyst fluid and PGD using embryo culture medium.

Objective: The aim of this study is to determine the sensitivity of the polymerase chain reaction (PCR) technique to detect the Y chromosome using cell-free DNA within a culture medium for gender prediction of blastocysts.

Materials And Methods: In this study, the gender of 30 embryos on day 5 was determined using embryonic DNA extraction from the culture medium and the PCR technique to evaluate the sex-determining region Y and fragile X mental retardation genes. Then, the accuracy was assessed using ultrasound.

Results: The results of the PCR technique showed that 7 embryos were male, but an ultrasound revealed that 13 were male.

Conclusion: The given results indicated that, because of the low amount of DNA extracted from the culture medium, the diagnosis of the existence of the Y chromosome by this method is still not accurate enough for detecting the gender of the embryo.

References

Galluzzi L, Palini S, De Stefani S, Andreoni F, Primiterra M, Diotallevi A . Extracellular embryo genomic DNA and its potential for genotyping applications. Future Sci OA. 2016; 1(4):FSO62. PMC: 5137924. DOI: 10.4155/fso.15.62. View

Handyside A, Kontogianni E, Hardy K, Winston R . Pregnancies from biopsied human preimplantation embryos sexed by Y-specific DNA amplification. Nature. 1990; 344(6268):768-70. DOI: 10.1038/344768a0. View

Li P, Song Z, Yao Y, Huang T, Mao R, Huang J . Preimplantation Genetic Screening with Spent Culture Medium/Blastocoel Fluid for in Vitro Fertilization. Sci Rep. 2018; 8(1):9275. PMC: 6006313. DOI: 10.1038/s41598-018-27367-4. View

Tasca R, McClure M . The emerging technology and application of preimplantation genetic diagnosis. J Law Med Ethics. 2000; 26(1):7-16, 3. DOI: 10.1111/j.1748-720x.1998.tb01901.x. View

Rajan-Babu I, Lian M, Cheah F, Chen M, Tan A, Prasath E . FMR1 CGG repeat expansion mutation detection and linked haplotype analysis for reliable and accurate preimplantation genetic diagnosis of fragile X syndrome. Expert Rev Mol Med. 2017; 19:e10. PMC: 5733830. DOI: 10.1017/erm.2017.10. View

Petersen C, Mauri A, Vagnini L, Renzi A, Petersen B, Matilla M . Randomized comparison of two commercial culture media (Cook and Vitrolife) for embryo culture after IMSI. JBRA Assist Reprod. 2018; 23(1):33-36. PMC: 6364275. DOI: 10.5935/1518-0557.20180058. View

Kuznyetsov V, Madjunkova S, Antes R, Abramov R, Motamedi G, Ibarrientos Z . Evaluation of a novel non-invasive preimplantation genetic screening approach. PLoS One. 2018; 13(5):e0197262. PMC: 5944986. DOI: 10.1371/journal.pone.0197262. View

Vera-Rodriguez M, Diez-Juan A, Jimenez-Almazan J, Martinez S, Navarro R, Peinado V . Origin and composition of cell-free DNA in spent medium from human embryo culture during preimplantation development. Hum Reprod. 2018; 33(4):745-756. DOI: 10.1093/humrep/dey028. View

Joshi B, De R, Goyal S . Utility and Applicability of a Universal Set of Primers in Identifying the Sex of South and Southeast Asian Mammals. Zool Stud. 2020; 58:e19. PMC: 6838184. DOI: 10.6620/ZS.2019.58-19. View

10.

Yang L, Lv Q, Chen W, Sun J, Wu Y, Wang Y . Presence of embryonic DNA in culture medium. Oncotarget. 2017; 8(40):67805-67809. PMC: 5620213. DOI: 10.18632/oncotarget.18852. View

11.

Meseguer M, Garrido N, Remohi J, Simon C, Pellicer A . Gender selection: ethical, scientific, legal, and practical issues. J Assist Reprod Genet. 2002; 19(9):443-6. PMC: 3455550. DOI: 10.1023/a:1016876026359. View

12.

Handyside A, Pattinson J, Penketh R, Delhanty J, Winston R, Tuddenham E . Biopsy of human preimplantation embryos and sexing by DNA amplification. Lancet. 1989; 1(8634):347-9. DOI: 10.1016/s0140-6736(89)91723-6. View

13.

Harper J, SenGupta S . Preimplantation genetic diagnosis: state of the art 2011. Hum Genet. 2011; 131(2):175-86. DOI: 10.1007/s00439-011-1056-z. View

14.

. Sex selection and preimplantation genetic diagnosis. Fertil Steril. 2004; 82 Suppl 1:S245-8. DOI: 10.1016/j.fertnstert.2004.05.016. View

15.

Basille C, Frydman R, El Aly A, Hesters L, Fanchin R, Tachdjian G . Preimplantation genetic diagnosis: state of the art. Eur J Obstet Gynecol Reprod Biol. 2009; 145(1):9-13. DOI: 10.1016/j.ejogrb.2009.04.004. View

16.

Handyside A, Lesko J, Tarin J, Winston R, Hughes M . Birth of a normal girl after in vitro fertilization and preimplantation diagnostic testing for cystic fibrosis. N Engl J Med. 1992; 327(13):905-9. DOI: 10.1056/NEJM199209243271301. View

17.

Settin A, Elsobky E, Hammad A, Al-Erany A . Rapid sex determination using PCR technique compared to classic cytogenetics. Int J Health Sci (Qassim). 2011; 2(1):49-52. PMC: 3068716. View

18.

Wu H, Ding C, Shen X, Wang J, Li R, Cai B . Medium-based noninvasive preimplantation genetic diagnosis for human α-thalassemias-SEA. Medicine (Baltimore). 2015; 94(12):e669. PMC: 4554004. DOI: 10.1097/MD.0000000000000669. View