Kyle J Tobler1, Yulian Zhao2, Ric Ross3, Andy T Benner4, Xin Xu4, Luke Du4, Kathleen Broman2, Kim Thrift2, Paul R Brezina5, William G Kearns6. 1. Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland; Department of Obstetrics and Gynecology, Womack Army Medical Center, Fort Bragg, North Carolina. Electronic address: kyle.tobler@gmail.com. 2. Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland. 3. Fort Worth Fertility, Fort Worth, Texas. 4. AdvaGenix, LLC, Rockville, Maryland. 5. Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Vanderbilt University School of Medicine, Memphis, Tennessee; Department of Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee; Fertility Associates of Memphis, Memphis, Tennessee. 6. Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland; AdvaGenix, LLC, Rockville, Maryland.
Abstract
OBJECTIVE: To obtain embryonic molecular karyotypes from genomic DNA (deoxyribonucleic acid) isolated from blastocoel fluid (BF) and to compare these karyotypes with the karyotypes from the remaining inner cell mass (ICM) and trophectoderm (TE) of the blastocyst. DESIGN: Prospective cohort study. SETTING: Academic center and preimplantation genetics laboratory. PATIENT(S): Ninety-six donated cryopreserved embryos. INTERVENTION(S): Embryo biopsy, BF aspiration, DNA analysis using a comparative genomic hybridization microarray (aCGH). MAIN OUTCOME MEASURE(S): The aCGH of a single blastomere, BF-DNA, and ICM-TE. RESULT(S): The BF-DNA samples resulted in a successful aCGH in 63% of cases. Discordance in karyotypes was found between the BF-DNA and the ICM-TE in 52% of cases. A total of 70% of aneusomic (mosaicism), cleavage-stage embryos differentiated into euploid blastocysts. Probabilities for diagnostic accuracy were calculated and demonstrated the following: sensitivity of 0.88 (95% confidence interval [CI]: 0.62-0.98); specificity of 0.55 (95% CI: 0.39-0.70); positive predictive value of 0.41 (95% CI: 0.25-0.60); negative predictive value of 0.92 (95% CI: 0.75-0.99). CONCLUSION(S): Genomic DNA from the BF can be amplified and characterized by comprehensive chromosome microarrays. The results demonstrated that aneusomic cleavage-stage embryos differentiated into euploid blastocysts, possibly using a mechanism that marginalizes aneuploid nuclei into the blastocoel cavity. In addition, owing to the high discordance between the karyotypes obtained from the BF-DNA and the ICM-TE, using BF-DNA for preimplantation genetic testing is not yet advised. Published by Elsevier Inc.
OBJECTIVE: To obtain embryonic molecular karyotypes from genomic DNA (deoxyribonucleic acid) isolated from blastocoel fluid (BF) and to compare these karyotypes with the karyotypes from the remaining inner cell mass (ICM) and trophectoderm (TE) of the blastocyst. DESIGN: Prospective cohort study. SETTING: Academic center and preimplantation genetics laboratory. PATIENT(S): Ninety-six donated cryopreserved embryos. INTERVENTION(S): Embryo biopsy, BF aspiration, DNA analysis using a comparative genomic hybridization microarray (aCGH). MAIN OUTCOME MEASURE(S): The aCGH of a single blastomere, BF-DNA, and ICM-TE. RESULT(S): The BF-DNA samples resulted in a successful aCGH in 63% of cases. Discordance in karyotypes was found between the BF-DNA and the ICM-TE in 52% of cases. A total of 70% of aneusomic (mosaicism), cleavage-stage embryos differentiated into euploid blastocysts. Probabilities for diagnostic accuracy were calculated and demonstrated the following: sensitivity of 0.88 (95% confidence interval [CI]: 0.62-0.98); specificity of 0.55 (95% CI: 0.39-0.70); positive predictive value of 0.41 (95% CI: 0.25-0.60); negative predictive value of 0.92 (95% CI: 0.75-0.99). CONCLUSION(S): Genomic DNA from the BF can be amplified and characterized by comprehensive chromosome microarrays. The results demonstrated that aneusomic cleavage-stage embryos differentiated into euploid blastocysts, possibly using a mechanism that marginalizes aneuploid nuclei into the blastocoel cavity. In addition, owing to the high discordance between the karyotypes obtained from the BF-DNA and the ICM-TE, using BF-DNA for preimplantation genetic testing is not yet advised. Published by Elsevier Inc.
Authors: Karen Sermon; Antonio Capalbo; Jacques Cohen; Edith Coonen; Martine De Rycke; Anick De Vos; Joy Delhanty; Francesco Fiorentino; Norbert Gleicher; Georg Griesinger; Jamie Grifo; Alan Handyside; Joyce Harper; Georgia Kokkali; Sebastiaan Mastenbroek; David Meldrum; Marcos Meseguer; Markus Montag; Santiago Munné; Laura Rienzi; Carmen Rubio; Katherine Scott; Richard Scott; Carlos Simon; Jason Swain; Nathan Treff; Filippo Ubaldi; Rita Vassena; Joris Robert Vermeesch; Willem Verpoest; Dagan Wells; Joep Geraedts Journal: Mol Hum Reprod Date: 2016-06-02 Impact factor: 4.025
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