Number of embryos from each cycle and the number of chromosomes assessed along with abnormalities detected.

<p>Rec: reciprocal translocation; Rob: Robertsonian translocation; Inv: Inversion; Bla: Blastomere; PB: polar body; TE: trophectoderm cells.</p>*<p>Chromosomes involved in the rearrangement were not assessed;</p>**<p>Abnormalities affecting chromosomes involved in the rearrangement were excluded. Case 12 was the only one in which two rearrangements were present. This case was excluded from the statistical analysis.</p

Malsegregation events detected in non-rearranged chromosomes in patient and control samples.

<p>Malsegregation events detected in non-rearranged chromosomes in patient and control samples.</p

Corrected number of errors in the patient-specific control groups.

*<p>Chromosomes involved in the matching patient's rearrangement were excluded from the assessed chromosomes.</p>**<p>Abnormalities affecting chromosomes involved in the matching patient's rearrangement were excluded.</p>***<p>Expected number in the control group if it consisted of the same number of samples as provided by the matched patient.</p

Patient information.

<p>Bla: Blastomere; PB: polar body; TE: trophectoderm cells; aCGH: Microarray-CGH; Met-CGH: Metaphase CGH. Note: Some patients underwent more than one cycle of PGD and may have had analysis conducted at different stages in subsequent cycles.</p

Data concerning the patient-matched control groups.

<p>Bla: Blastomere; PB: polar body; TE: trophectoderm cells;</p>*<p>Chromosomes involved in the matching patient's rearrangement were excluded from the assessed chromosomes.</p>**<p>Abnormalities affecting chromosomes involved in the matching patient's rearrangement were excluded.</p

Microarray-CGH analysis of an embryo from a Robertsonian translocation carrier.

<p>Cytogenetic analysis of a cleavage stage embryo from a Robertsonian translocation carrier- 46,XY,t(13;15)(q21.3;q11.2). Microarray-CGH revealed monosomy 13, presumably resulting from a meiotic error due to problems processing the rearranged chromosomes. An additional aneuploidy unrelated to the Robertsonian translocation (monosomy for chromosome 1) was also detected. The two monosomies are indicated by the altered ratio of fluorescence related to the test (embryo) and reference (46,XY) DNA samples. All of the probes corresponding to chromosomes 1 and 13 have test/reference ratios less than 0.3.</p

mtDNA quantities and clinical outcomes of 23 TE samples assessed via real-time PCR and NGS.

<p>*The threshold for considering a sample to have elevated mtDNA levels, incompatible with implantation, was 0.003 for real-time PCR and 0.07 for NGS.</p><p>mtDNA quantities and clinical outcomes of 23 TE samples assessed via real-time PCR and NGS.</p

The relationship between mtDNA quantity, female age and embryo chromosome constitution.

<p><b>a)</b> Data obtained during quantitative real-time PCR analysis of TE samples removed from 302 blastocysts demonstrated a statistically significant increase (P = 0.003) in the level of mtDNA in relation to advancing female age. This phenomenon was evident for both euploid and aneuploid blastocysts. <b>b)</b> Real-time PCR analysis of 39 blastomeres showed that cleavage stage embryos from reproductively younger women contained significantly (P = 0.01) higher mtDNA levels, compared to those generated by reproductively older women. <b>c)</b> Real-time PCR analysis of TE samples also demonstrated that aneuploid blastocysts (n = 99) contained significantly (P = 0.025) larger quantities of mtDNA at all ages, compared to those that were euploid (n = 203). Statistical analysis of mtDNA values took place with the use of unpaired two-tailed t-tests.</p

Blastocyst mtDNA quantity threshold in relation to clinical outcome.

<p><b>a)</b> The mtDNA quantity viability threshold for euploid blastocysts, established via retrospective analysis of TE biopsies from transferred embryos with known outcomes. All blastocysts producing viable pregnancies contained mtDNA quantities below the 0.003 value (red line) whereas mtDNA quantities above this value were associated with failure to achieve an ongoing clinical pregnancy. <b>b)</b> Results of the prospective blinded study. The mtDNA threshold used was the same as that established in the retrospective study (4a). Validity was confirmed, since all blastocysts producing viable pregnancies contained mtDNA quantities below the cut-off (red line) and no blastocysts with mtDNA quantities above this value achieved an ongoing clinical pregnancy. <b>c)</b> NGS analysis of the mtDNA level in 23 euploid TE samples. The corresponding embryos were transferred during SET cycles, and clinical outcomes were known for 21 of them. As with the real-time PCR experiments, mtDNA levels were lower in the seven implanting embryos (note- the y-axis scale is different for NGS analyses and consequently cut-off values differ).</p

The average relative quantities of mtDNA observed in association with female age at the cleavage stage.

<p>The mtDNA values (2-^{Delta Delta Ct}) were obtained during real-time PCR analysis. All examined blastomeres were characterised as being chromosomally normal.</p><p>The average relative quantities of mtDNA observed in association with female age at the cleavage stage.</p