Screening method of different metabolites for evaluating goat transgenic cloned embryo quality based on gas chromatography-mass spectrum combined technique and morphology

Abstract

The invention discloses a screening method of different metabolites for evaluating goat transgenic cloned embryo quality based on a gas chromatography-mass spectrum combined technique and morphology. The screening method comprises the following steps that 1, reconstructed embryos are cultured for 48 hours, the cleavage rate is checked, cleaved embryos are divided into a high-quality cleaved embryo group and a low-quality cleaved embryo group according to the morphology, the embryo groups are randomly put into blastocyst culture droplets for continuous culture; 2, the blastocyst rate statistics is performed at 7.5 d, and blastocysts and embryo metabolites of the high-quality cleaved embryo group and the low-quality cleaved embryo group are respectively collected; 3, the collected embryo metabolite samples are processed, and gas chromatography-mass spectrum detection is performed; 4, detection results are subjected to statistic analysis. The screening method is integrated with morphological evaluation of the embryos to find metabolism markers closely correlated with the goat transgenic cloned embryo quality, provides a reference for research on prediction of embryonic developmental potential and facilitates screening of high-quality transplanted embryos and improvement of transgenic cloning efficiency.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method for screening differential metabolites of goat transgenic cloned embryo quality assessment based on gas chromatography-mass spectrometry technology and morphology. Background technique [0002] Somatic cell nuclear transfer (SCNT) can dedifferentiate somatic cells in vitro and develop into totipotent embryos, which has broad clinical application prospects. However, the process of SCNT technology is complicated and there are many influencing factors, and the efficiency of obtaining newborn individuals through SCNT technology is also very low, which seriously restricts the research and application of transgenic large livestock. The study found that the lack of an effective embryo evaluation system and the inability to screen out high-quality embryos for transfer are one of the main reasons for the low efficiency of SCNT. Therefore, it is very important to find a ...

AI Technical Summary

Problems solved by technology

Although this method is intuitive and fast, it lacks a unified standard, and the subjectivity of the observer makes the assessment of embryo morphology and developmental potential biased.

Alternatively, embryos with embryo fragments may have better developmental potential, and embryos with normal morphology may also have genetic or epigenetic defects

Therefore, morphology alone is not sufficient to identify high-quality embryos

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