P15 PGD by FISH in India: first live-birth after PGD for a reciprocal translocation

Abstract

expression pattern of specific genes important for fetal growth and nutrition. Materials and Methods: Blood samples from children conceived after ART (n = 54) and from spontaneously conceived children (n = 29) were collected after informed consent in PAXGene Blood RNA Tubes. The children were part of an existing cohort from the UK, followed since birth. Blood samples were stored at 20oC and transferred frozen to Belgium. RNA was extracted for reverse transcription and Real-Time PCR analysis using gene expression assays for DNA methyltransferase 1 (DNMT1), Glucocorticoid receptor (NR3C1) and Peroxisome Proliferator Activated Receptor Alpha (PPARA) and Hypoxanthine Phosphoribosyltransferase 1 (HPRT1) as an endogenous control. Statistical analysis was done using the Student T test. Results: Gene expression levels were normalised against the endogenous control HPRT1 and relatively quantified using the DDCt method. The mean blood mRNA levels of DNMT1, NR3C1 and PPARA in the ART children show a 0.95, 1.26 and 1.35 fold change respectively compared to those of the children from spontaneous conception (p = 0.95, p = 0.73 and p = 0.54 respectively). Discussion and Conclusion: No significant difference was observed in mRNA expression levels of DNMT1, NR3C1 and PPARA between children conceived after ART and children spontaneously conceived, reassuring the epigenetic safety of ART methods with respect to the studied genes. Nevertheless, no definitive conclusions can be drawn without testing other gene panels of imprinted and non-imprinted genes.