Raman spectroscopy analysis of differences in composition of spent culture media of in vitro cultured preimplantation embryos isolated from normal and fat mice dams.
The aim of the present study was to compare overall patterns of metabolic activity of in vitro cultured preimplantation embryos isolated from normal and fat mice dams by means of non-invasive profiling of spent culture media using Raman spectroscopy. To produce females with two different types of body condition (normal and fat), a previously established two-generation model was used, based on overfeeding of experimental mice during prenatal and early postnatal development. Embryos were isolated from spontaneously ovulating and naturally fertilized dams at the 2-cell stage of development and cultured to the blastocyst stage in synthetic oviductal medium KSOMaa. Embryos from fat mice (displaying significantly elevated body weight and fat) showed similar developmental capabilities in vitro as embryos isolated from normal control dams (displaying physiological body weight and fat). The results show that alterations in the composition of culture medium caused by the presence of developing mouse preimplantation embryos can be detected using Raman spectroscopy. Metabolic activity of embryos was reflected in evident changes in numerous band intensities in the 1620-1690cm(-1) (amide I) region and in the 1020-1140cm(-1) region of the Raman spectrum for KSOMaa. Moreover, multivariate analysis of spectral data proved that the composition of proteins and other organic compounds in spent samples obtained after the culture of embryos isolated from fat dams was different from that in spent samples obtained after the culture of embryos from control dams. This study demonstrates that metabolic activity of cultured preimplantation embryos might depend on the body condition of their donors.
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, 04012 Košice, Slovakia. Electronic address: fabian@saske.sk.
Dušan
D
Fabian
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, 04012 Košice, Slovakia.
Martina
M
Kačmarová
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, 04012 Košice, Slovakia.
Janka
J
Kubandová
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, 04012 Košice, Slovakia.
Štefan
Š
Čikoš
Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, 04012 Košice, Slovakia.
Juraj
J
Koppel
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D003470:Culture Media / Q000737:chemistry*; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro; D051379:Mice; D009765:Obesity / Q000378:metabolism*; D013059:Spectrum Analysis, Raman
D003470:Culture Media
In vitro culture; Obesity; Preimplantation embryo; Raman spectroscopy