Centre for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital; National Clinical Research Center for Obstetrics and Gynecology(Peking University Third Hospital); Ministry of Education, Key Laboratory of Assisted Reproduction (Peking University); Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology.
Department of Clinical Research, Yikon Genomics Co. Ltd.
J Vis Exp. 2021 Sep 7(175). doi: 10.3791/62619.
In clinical in vitro fertilization (IVF), the prevailing method for PGT-A requires biopsy of a few cells from the trophectoderm (TE). This is the lineage that forms the placenta. This method, however, requires specialized skills, is invasive, and suffers from false positives and negatives because the chromosome numbers in the TE and the inner cell mass (ICM), which develops into the fetus, are not always the same. NICS, a technology requiring sequencing of DNA that released into the culture medium from both TE and ICM, may offer a way out to these problems but has previously been shown to have limited efficacy. The present study reports the full protocol of NICS, which includes culture medium sampling methods, whole genome amplification (WGA) and library preparation, and NGS data analysis by analysis software. Considering the different cryopreservation times in different embryo laboratories, embryologists have two methods for collecting embryo culture medium that can be selected according to the actual conditions of the IVF laboratory.
在临床体外受精 (IVF) 中，PGT-A 的主要方法需要从滋养外胚层 (TE) 中活检少数细胞。滋养外胚层是形成胎盘的谱系。然而，这种方法需要专门的技能，具有侵入性，并且存在假阳性和假阴性，因为滋养外胚层和内细胞团 (ICM) 中的染色体数量并不总是相同的，内细胞团会发育成胎儿。NICS 是一种需要测序 DNA 的技术，从 TE 和 ICM 释放到培养基中，这可能为解决这些问题提供了一种方法，但之前已显示其疗效有限。本研究报告了 NICS 的完整方案，包括培养基采样方法、全基因组扩增 (WGA) 和文库制备，以及通过分析软件进行 NGS 数据分析。考虑到不同胚胎实验室的不同冷冻保存时间，胚胎学家有两种收集胚胎培养基的方法，可以根据 IVF 实验室的实际情况选择。
