Investigation into the Use of Nuclear Magnetic Resonance Spectroscopy for Viability Predictions, in Real Time, on Mouse Preimplantation Embryos

Joseph T. Lyman, Marcello Rubessa, Deborah K. Oh, Ginger L. Torres, Richard L. Magin ,Matthew B. Wheeler

In Western societies, up to 4% of children born are conceived using assisted reproduction technologies (ARTs), with IVF (in vitro fertilization) now the clinically preferred method of treatment for infertility. It is known that nuclear magnetic resonance (NMR) spectroscopy can be used to identify and quantify metabolite levels in cells and embryos. In these experiments we evaluated embryo metabolism in real time. The development of preimplantation mouse embryos subjected to NMR was compared to embryos subjected only to the manipulations (SHAM), to embryos subjected to deuterium-containing medium (D₂O), and to control embryos. Mouse embryos analyzed with NMR, at the two-cell stage, exhibited lower (p<0.05) development to the blastocyst stage than control embryos.  SHAM treated and NMR analyzed embryos developed at higher rates than embryos only exposed to D₂0 at 37 ºC, indicating attenuation of D₂O effects at lower temperatures. To determine why NMR analysis attenuates effects of exposure to D₂O, embryos were cultured to blastocysts after exposure to D₂O at ambient or 37 ºC.  Results showed that at lower temperatures, exposure to D₂O was less harmful. Finally, transfer of embryos into recipients after NMR resulted in two litters of normal pups. This study suggests that NMR techniques can be used for embryo evaluation.