Preliminary Study On Establishment Of A Prediction Model For The Developmental Potential Of Early Buffalo Embryo Based On Metabonomics

The dynamical changes of embryonic metabolism, which reflect the embryonic developmental state and response to internal and external environment, can be used as evaluation criteria of the embryonic development potential. The effect of overall prediction model based on embryonic metabolism can get better consistency and predictability than that of the traditional evaluation method based on the morphology of embryo. The present study was to establish an individual buffalo embryo culture system, and use a classifier to separate difference buffalo embryo potentialities of development by use the cultured medium of Raman spectroscopy fingerprint data. The methods and results of the experiments were summarized as follows:1. To preliminarily establish a two-step culture system with SOF medium by improved our conventional embryo culture system. The result showed that even though the feeder and serum free culture system had a significantly lower blastocyst formation rate from morula-stage to blastocyst compared to the group culture system with feeder cells and serum (P<0.05,51.2%,63/123VS.40.4%,59/146) it could support individual buffalo embryo development from morula-stage to blastocyst when SOF medium was used.2. To investigate the variation of Raman spectrum in serum and BSA culture system. The results showed that background differences of Raman spectrum in serum culture system showed more obvious than in BSA culture system. These data showed that the non-serum culture system was advantage on detecting Raman spectrum of the buffalo embryo culture medium for subsequently establishing a developmental potential model.3. A support vector machine was adopted by parameter optimization with different methods to analyze the Raman spectrum of buffalo’s embryonic culture medium, and obtained the reliable prediction results: Classification accuracy98.6%and prediction accuracy100%.In conclusion, an individual bufflao embryo culuture was preliminarily established by improved our conventional embryo culture system, meanwhile, the effects on variation of Raman spectrum in serum and BSA culture system were detected; a support vector machine was adopted by parameter optimization with different methods to analyze the Raman spectrum of buffalo’s embryonic culture medium, and obtained the reliable prediction results:Classification accuracy98.6%and prediction accuracy100%. Base on a new overall viewpoint to investigate the dynamic development of buffalo’s embryo and the characteristic of Raman instrument, the established model for predicting the developmental potential of buffalo embryo in this study is a complement and development of the traditional evaluation system, which has obvious advantages on overall objective, rapid, accurate and non-invasive compared with traditional evaluation system.