FT-IR spectroscopic metabolic fingerprinting of human embryo culture media combined with multivariate statistical analysis to discriminate embryos with different reproductive potential

M. GARCEA; ALEJANDRA BOSCH; M. GOMEZ PEÑA; PABLO MARTINA; OSVALDO YANTORNO; C. BISIOLI.

Denver, Colorado

Congreso; 66th Annual Meeting, American Society for Reproductive Medicine,; 2010

American Society for Reproductive Medicine

FT-IR spectroscopy of biological materials provides highly specific fingerprint-like signatures of their structural composition. The aim of this work was to study metabolic fingerprinting of spent culture media of human embryos and to develop a multivariate statistical model to discriminate embryonic reproductive potential. MATERIALS AND METHODS We analyzed 30 spent day 3-embryo culture media from 15 patients with known outcome after IVF-ET, using FT-IR spectroscopy. 30 ƒÊL of embryo culture media were transferred to ZnSe optical plates and dried to obtain transparent films. FT-IR spectra were acquired between 4000 and 650 cm-1 with 6 cm-1 spectral resolution using a Spectrum One FT-IR spectrometer. To increase the number of discriminative features present in spectra, the first and second derivatives were calculated using OPUS software (Bruker Optics). Dendrograms were constructed by executing WardLs algoritm of OPUS software using first derivate spectra as input data for multivariate hierarchical cluttering analysis ( HCA). ƒÊL of embryo culture media were transferred to ZnSe optical plates and dried to obtain transparent films. FT-IR spectra were acquired between 4000 and 650 cm-1 with 6 cm-1 spectral resolution using a Spectrum One FT-IR spectrometer. To increase the number of discriminative features present in spectra, the first and second derivatives were calculated using OPUS software (Bruker Optics). Dendrograms were constructed by executing WardLs algoritm of OPUS software using first derivate spectra as input data for multivariate hierarchical cluttering analysis ( HCA). RESULTS We found that in vitro cultured embryos that resulted in pregnancy produce a different spectroscopic pattern compared with embryos that do not result in pregnancy. We could determine that these differences could be detectable in first derivate spectra in the 3000-2800 cm-1 range (lipid region), the 1500-1300 cm-1 interval (mixed region) and the 1200-900 cm-1 region (carbohydrate region).When HCA was applied to first derivate spectra in those spectral ranges, the dendrogram produced showed two clusters which clearly sep rate embryos with proven pregnancy from those which failed to implant. CONCLUSIONS: Our preliminary study shows that changes detected in spectroscopic metabolic fingerprinting of spent human culture media from embryos with different reproductive potential would allow the development of a robust, fast and noninvasive methodology based on a multivariate statistical analysis able to predict pregnancy outcome before embryo transfer . We found that in vitro cultured embryos that resulted in pregnancy produce a different spectroscopic pattern compared with embryos that do not result in pregnancy. We could determine that these differences could be detectable in first derivate spectra in the 3000-2800 cm-1 range (lipid region), the 1500-1300 cm-1 interval (mixed region) and the 1200-900 cm-1 region (carbohydrate region).When HCA was applied to first derivate spectra in those spectral ranges, the dendrogram produced showed two clusters which clearly sep rate embryos with proven pregnancy from those which failed to implant. CONCLUSIONS: Our preliminary study shows that changes detected in spectroscopic metabolic fingerprinting of spent human culture media from embryos with different reproductive potential would allow the development of a robust, fast and noninvasive methodology based on a multivariate statistical analysis able to predict pregnancy outcome before embryo transfer . CONCLUSIONS: Our preliminary study shows that changes detected in spectroscopic metabolic fingerprinting of spent human culture media from embryos with different reproductive potential would allow the development of a robust, fast and noninvasive methodology based on a multivariate statistical analysis able to predict pregnancy outcome before embryo transfer .