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回到未来:优化的单个人类胚胎植入前阶段的微滴培养。

Back to the future: optimised microwell culture of individual human preimplantation stage embryos.

机构信息

RVT Australia, Cairns, QLD, 4870, Australia.

VitaVitro Biotech Co., Ltd., Shenzhen, China.

出版信息

J Assist Reprod Genet. 2021 Oct;38(10):2563-2574. doi: 10.1007/s10815-021-02167-4. Epub 2021 Apr 16.

DOI:10.1007/s10815-021-02167-4
PMID:33864207
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8581087/
Abstract

Although in vitro culture of human embryos is a crucial step in assisted reproduction, the lack of focused research hampers worldwide standardisation and consistent outcomes. Only 1.2% of research papers published in five leading journals in human reproduction in 2019 focused on in vitro culture conditions, creating the impression that the optimisation process has approached its limits. On the other hand, in vitro culture of mammalian embryos is based on old principles, while there is no consensus on basic issues as density, time, medium change, gas atmosphere and small technical details including the way of drop preparation. This opinion paper aims to highlight and analyse the slow advancement in this field and stimulate research for simple and affordable solutions to meet the current requirements. A possible way for advancement is discussed in detail. Selection of embryos with the highest developmental competence requires individual culture and modification of the widely used "drop under oil" approach. Current use of three-dimensional surfaces instead of large flat bottoms is restricted to time-lapse systems, but these wells are designed for optical clarity, not for the needs of embryos. The size and shape of the original microwells (Well of the Well; WOW) offer a practical and straightforward solution to combine the benefits of communal and individual incubation and improve the overall quality of cultured embryos.

摘要

尽管体外培养人类胚胎是辅助生殖的关键步骤,但缺乏重点研究阻碍了全球标准化和结果的一致性。2019 年,在人类生殖领域的五本领先期刊上发表的研究论文中,仅有 1.2%的论文聚焦于体外培养条件,给人一种优化过程已经接近极限的印象。另一方面,哺乳动物胚胎的体外培养基于旧的原则,而在基本问题上,如密度、时间、培养基更换、气体环境以及包括滴液制备方式在内的微小技术细节上,都没有达成共识。本文旨在强调和分析该领域进展缓慢的问题,并为满足当前需求,推动简单且经济实惠的解决方案的研究。详细讨论了一种可能的进展方式。选择具有最高发育能力的胚胎需要进行单独培养,并对广泛使用的“油下滴液”方法进行修改。目前,三维表面的使用代替了大型平底,仅限于延时拍摄系统,但这些孔是为光学清晰度而设计的,而不是为胚胎的需求而设计的。原始微腔(Well of the Well;WOW)的大小和形状提供了一个实用且直接的解决方案,可结合集体和个体孵育的优势,提高培养胚胎的整体质量。

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In Vitro Culture of Mammalian Embryos: Is There Room for Improvement?哺乳动物胚胎的体外培养:是否有改进的空间?
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Individually Cultured Bovine Zygotes Successfully Develop to the Blastocyst Stage in an Extremely Confined Environment.在极其受限的环境中,个体培养的牛胚胎成功发育到囊胚阶段。
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本文引用的文献

1
Is cell-free DNA in spent embryo culture medium an alternative to embryo biopsy for preimplantation genetic testing? A systematic review.胚胎培养液中的游离细胞 DNA 是否可替代胚胎活检用于植入前遗传学检测?系统综述。
Reprod Biomed Online. 2020 Jun;40(6):779-796. doi: 10.1016/j.rbmo.2020.02.002. Epub 2020 Feb 20.
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Vitrification in ART: past, present, and future.ART 中的玻璃化:过去、现在和未来。
Theriogenology. 2020 Jul 1;150:276-279. doi: 10.1016/j.theriogenology.2020.01.057. Epub 2020 Feb 19.
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Need for choosing the ideal pH value for IVF culture media.需要选择理想的 pH 值用于 IVF 培养液。
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The impact of selected embryo culture conditions on ART treatment cycle outcomes: a UK national study.特定胚胎培养条件对辅助生殖技术治疗周期结局的影响:一项英国全国性研究。
Hum Reprod Open. 2020 Feb 10;2020(1):hoz031. doi: 10.1093/hropen/hoz031. eCollection 2020.
5
Effect of Group Embryo Culture under Low-Oxygen Tension in Benchtop Incubators on Human Embryo Culture: Prospective, Randomized, Controlled Trial.低氧张力台式培养箱中分组培养对人类胚胎培养的影响:前瞻性、随机、对照试验。
Reprod Sci. 2020 Jul;27(7):1522-1533. doi: 10.1007/s43032-020-00150-5. Epub 2020 Jan 28.
6
'There is only one thing that is truly important in an IVF laboratory: everything' Cairo Consensus Guidelines on IVF Culture Conditions.《体外受精实验室唯一真正重要的事情:一切》开罗体外受精培养条件共识指南。
Reprod Biomed Online. 2020 Jan;40(1):33-60. doi: 10.1016/j.rbmo.2019.10.003. Epub 2019 Oct 10.
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Temperature of embryo culture for assisted reproduction.辅助生殖中胚胎培养的温度。
Cochrane Database Syst Rev. 2019 Sep 17;9(9):CD012192. doi: 10.1002/14651858.CD012192.pub2.
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Unstable osmolality of microdrops cultured in non-humidified incubators.非加湿培养箱中培养的微滴渗透压不稳定。
J Assist Reprod Genet. 2019 Aug;36(8):1571-1577. doi: 10.1007/s10815-019-01515-9. Epub 2019 Jul 2.
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Culture of Human Preimplantation Embryos in a Clinical ART Setting.临床辅助生殖技术环境下人类植入前胚胎的培养
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Controversies in ART: considerations and risks for uninterrupted embryo culture.ART 相关争议:胚胎连续培养的考虑因素和风险
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