Embryo Culture Techniques

胚胎培养技术
  • 文章类型: Guideline
    本会议报告介绍了国际专家会议的结果,以建立有关IVF培养条件的共识指南。审查和讨论的主题是:胚胎培养-基本原理和相互作用;IVF实验室的温度;培养中的湿度;二氧化碳控制和中等pH值;胚胎培养的氧气张力;工作站-设计和工程;孵化器-维持培养环境;显微操作-维持稳定的物理化学环境;处理实践;评估实践;培养基-缓冲和pH值,一般成分和蛋白质补充剂,用于人类胚胎培养的顺序或单步培养基;使用和管理-冷链和储存;测试设备-校准和认证;以及实验室设备和实时监控。在这些一般标题下建立了50多个共识准则点。
    This proceedings report presents the outcomes from an international expert meeting to establish consensus guidelines on IVF culture conditions. Topics reviewed and discussed were: embryo culture - basic principles and interactions; temperature in the IVF laboratory; humidity in culture; carbon dioxide control and medium pH; oxygen tension for embryo culture; workstations - design and engineering; incubators - maintaining the culture environment; micromanipulation - maintaining a steady physcochemical environment; handling practices; assessment practices; culture media - buffering and pH, general composition and protein supplementation, sequential or single-step media for human embryo culture; use and management - cold chain and storage; test equipment - calibration and certification; and laboratory equipment and real-time monitoring. More than 50 consensus guideline points were established under these general headings.
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  • 文章类型: Journal Article
    目的:确定速裂D3胚胎的发育能力。
    方法:回顾性研究包括2014年7月至2017年6月进行的513个PGT-A周期的4028个胚胎。胚胎在延时培养箱中培养,并在囊胚期进行活检。根据D3上的细胞数量(从2个细胞到≥13个细胞并压实)将胚胎分类。进行了针对混杂因素进行调整的广义线性混合模型,以评估与8细胞胚胎的机会相比,每组中产生整倍体胚泡的机会。还分析了着床率和活产率。
    结果:统计分析表明,具有9至11个细胞的胚胎的整倍体囊胚率略低于8个细胞的胚胎(OR(95%CI)0.77(0.61-0.96)),而发现具有11个以上细胞的胚胎与8个细胞的胚胎一样可能产生整倍体囊胚(OR(95%CI)1.56)(0.20)。相反,慢切割胚胎的整倍体囊胚率显著低于8细胞胚胎(OR(95%CI)0.31(0.24-0.39)).此外,来自快速切割胚胎和8细胞胚胎的整倍体胚泡表现出相似的活产率。在8细胞和9至11细胞胚胎(OR(95%CI)1.23(0.70-2.15))和>11细胞胚胎(OR(95%CI)1.09(0.57-2.09))之间,活产的机会没有显着差异。
    结论:具有超过11个细胞的胚胎表现出与8细胞胚胎相似的发育能力。应重新考虑其不良预后。
    OBJECTIVE: To determine the developmental competence of fast-cleaving D3 embryos.
    METHODS: Retrospective study including 4028 embryos from 513 PGT-A cycles performed between July 2014 and June 2017. Embryos were cultured in time-lapse incubators and biopsied at blastocyst stage. Embryos were classified in groups according to the number of cells on D3 (from 2-cell to ≥13 -cell and compacted). A generalized linear mixed model adjusted for confounding factors was performed to assess the chance to give rise to an euploid blastocyst in each group compared with the chance of 8-cell embryos. Implantation and live birth rates were also analyzed.
    RESULTS: The statistical analysis showed that embryos with 9 to 11 cells had a slightly lower euploid blastocyst rate than 8-cell embryos (OR (95% CI) 0.77 (0.61-0.96)) while embryos with more than 11 cells were found to be just as likely to give rise to an euploid blastocyst as the 8-cell embryos (OR (95% CI) 1.20 (0.92-1.56)). Conversely, slow-cleaving embryos had a significantly lower euploid blastocyst rate than 8-cell embryos (OR (95% CI) 0.31 (0.24-0.39)). Moreover, euploid blastocysts derived from fast-cleaving embryos and from 8-cell embryos exhibit similar live birth rates. No significant differences were found in the chance to give rise a live birth between 8-cell and 9- to 11-cell embryos (OR (95% CI) 1.23 (0.70-2.15)) and > 11-cell embryos (OR (95% CI) 1.09 (0.57-2.09)).
    CONCLUSIONS: Embryos with more than 11 cells exhibit similar developmental competence to 8-cell embryos. Their poor prognosis should be reconsidered.
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  • 文章类型: Editorial
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  • 文章类型: Journal Article
    OBJECTIVE: Which recommendations can be provided by the European Society of Human Reproduction and Embryology Special Interest Group (ESHRE SIG) Embryology to support laboratory specialists in the organization and management of IVF laboratories and the optimization of IVF patient care?
    CONCLUSIONS: Structured in 13 sections, the guideline development group formulated recommendations for good practice in the organization and management of IVF laboratories, and for good practice of the specific procedures performed within the IVF laboratory.
    BACKGROUND: NA.
    METHODS: The guideline was produced by a group of 10 embryologists representing different European countries, settings and levels of expertise. The group evaluated the document of 2008, and based on this assessment, each group member rewrote one or more sections. Two 2-day meetings were organized during which each of the recommendations was discussed and rewritten until consensus within the guideline group was reached. After finalizing the draft, the members of the ESHRE SIG embryology were invited to review the guideline.
    METHODS: NA.
    RESULTS: The guideline provides recommendations on the general organization of an IVF laboratory (staffing and direction, quality management, laboratory safety), and on the specific aspects of the procedures performed in IVF laboratories (Identification of patients and traceability of their reproductive cells, consumables, handling of biological material, oocyte retrieval, sperm preparation, insemination of oocytes, scoring for fertilization, embryo culture and transfer, and cryopreservation). A last section provides recommendations regarding an Emergency plan for IVF laboratories.
    CONCLUSIONS: Evidence on most of the issues described is scarce, and therefore it was decided not to perform a formal search for and assessment of scientific evidence. However, recommendations published in the EUTCD and relevant and recent documents, manuals and consensus papers were taken into account when formulating the recommendations.
    CONCLUSIONS: Despite the limitations, the guideline group is confident that this document will be helpful to directors and managers involved in the management and organization of IVF laboratories, but also to embryologists and laboratory technicians performing daily tasks.
    BACKGROUND: The guideline was developed and funded by ESHRE, covering expenses associated with the guideline meetings. The guideline group members did not receive payment. Dr Coticchio reports speaker\'s fees from IBSA and Cook, outside the submitted work; Dr Lundin reports grants from Vitrolife, personal fees from Merck Serono, non-financial support from Unisense, outside the submitted work; Dr. Rienzi reports personal fees from Merck Serono, personal fees from MSD, grants from GFI, outside the submitted work; the other authors had nothing to disclose.
    BACKGROUND: NA.
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  • 文章类型: Journal Article
    OBJECTIVE: Can the approach to, and terminology for, time-lapse monitoring of preimplantation embryo development be uniformly defined in order to improve the utilization and impact of this novel technology?
    CONCLUSIONS: The adoption of the proposed guidelines for defining annotation practice and universal nomenclature would help unify time-lapse monitoring practice, allow validation of published embryo selection algorithms and facilitate progress in this field.
    BACKGROUND: An increasing quantity of publications and communications relating to time-lapse imaging of in vitro embryo development have demonstrated the added clinical value of morphokinetic data for embryo selection. Several articles have identified similar embryo selection or de-selection variables but have termed them differently. An evidence-based consensus document exists for static embryo grading and selection but, to date, no such reference document is available for time-lapse methodology or dynamic embryo grading and selection.
    METHODS: A series of meetings were held between September 2011 and May 2014 involving time-lapse users from seven different European centres. The group reached consensus on commonly identified and novel time-lapse variables.
    METHODS: Definitions, calculated variables and additional annotations for the dynamic monitoring of human preimplantation development were all documented.
    RESULTS: Guidelines are proposed for a standard methodology and terminology for the of use time-lapse monitoring of preimplantation embryo development.
    CONCLUSIONS: The time-lapse variables considered by this group may not be exhaustive. This is a relatively new clinical technology and it is likely that new variables will be introduced in time, requiring revised guidelines. A different group of users from those participating in this process may have yielded subtly different terms or definitions for some of the morphokinetic variables discussed. Due to the technical processes involved in time-lapse monitoring, and acquisition of images at varied intervals through limited focal planes, this technology does not currently allow continuous monitoring such that the entire process of preimplantation embryo development may be visualized.
    UNASSIGNED: This is the first time that a group of experienced time-lapse users has systematically evaluated current evidence and theoretical aspects of morphokinetic monitoring to propose guidelines for a standard methodology and terminology of its use and study, and its clinical application in IVF. The adoption of a more uniform approach to the terminology and definitions of morphokinetic variables within this developing field of clinical embryology would allow practitioners to benefit from improved interpretation of data and the sharing of best practice and experience, which could impact positively and more swiftly on patient treatment outcome.
    BACKGROUND: There was no specific funding for the preparation of these proposed guidelines. Meetings were held opportunistically during scientific conferences and using online communication tools. H.N.C. is a scientific consultant for ESCO, supplier of Miri TL. I.E.A. is a minor shareholder in Unisense Fertilitech, supplier of the EmbryoScope. Full disclosures of all participants are presented herein. The remaining authors have no conflict of interest.
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  • 文章类型: Journal Article
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  • 文章类型: Journal Article
    BACKGROUND: Recently we reported the development and optimization of a zebrafish teratogenicity assay using dechorionated AB strain embryos, a promising assay that was 87% concordant in correctly identifying in vivo teratogens and non-teratogens from a set of 31 compounds (Brannen et al., 2010: Birth Defects Res 89:66-77).
    METHODS: This assay utilizes a zebrafish morphological score system to characterize adverse effects and identify the no-observed-adverse-effect level (NOAEL).
    RESULTS: This report describes in detail the morphological score system used in the dechorionated zebrafish embryo culture teratogenicity assay. The morphological assessment includes evaluation of most structures and organ systems and grades relative severity of abnormalities.
    CONCLUSIONS: To this end, the morphological score system provides information of tissue-specific teratogenicity that has been found to have good concordance with structures found affected in vivo and can also be used to rank compounds based on the severity of malformations.
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