Reproductive technology revolutionized dairy production during the past century. Artificial insemination was first successfully applied to cattle in the early 1900s. The next major developments involved semen extenders, invention of the electroejaculator, progeny testing, addition of antibiotics to semen during the 1930s and 1940s, and the major discovery of sperm cryopreservation with glycerol in 1949. The 1950s and 1960s were particularly productive with the development of protocols for the superovulation of cattle with both pregnant mare serum gonadotrophin/equine chorionic gonadotrophin and FSH, the first successful bovine embryo transfer, the discovery of sperm capacitation, the birth of rabbits after in vitro fertilization, and the development of insulated liquid nitrogen tanks. Improved semen extenders and the replacement of glass ampules with plastic semen straws followed. Some of the most noteworthy developments in the 1970s included the initial successes with in vitro culture of embryos, calves born after chromosomal sexing as embryos, embryo splitting resulting in the birth of twins, and development of computer-assisted semen analysis. The 1980s brought flow cytometric separation of X- and Y-bearing sperm, in vitro fertilization leading to the birth of live calves, clones produced by nuclear transfer from embryonic cells, and ovum pick-up via ultrasound-guided follicular aspiration. The 20th century ended with the birth of calves produced from AI with sexed semen, sheep and cattle clones produced by nuclear transfer from adult somatic cell nuclei, and the birth of transgenic cloned calves. The 21st century has seen the introduction of perhaps the most powerful biotechnology since the development of artificial insemination and cryopreservation. Quick, inexpensive genomic analysis via the use of single nucleotide polymorphism genotyping chips is revolutionizing the cattle breeding industry. Now, with the introduction of genome editing technology, the changes are becoming almost too rapid to fully digest.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

A commercially viable cattle embryo transfer (ET) industry was established in North America during the early 1970s, approximately 80 years after the first successful embryo transfer was reported in a mammal. Initially, techniques for recovering and transferring cattle embryos were exclusively surgical. However, by the late 1970s, most embryos were recovered and transferred nonsurgically. Successful cryopreservation of embryos was widespread by the early 1980s, followed by the introduction of embryo splitting, in vitro procedures, direct transfer of frozen embryos and sexing of embryos. The wide spread adoption of ethylene glycol as a cryoprotectant has simplified the thaw-transfer procedures for frozen embryos. The number of embryos recovered annually has not grown appreciably over the last 10 years in North America and Europe; however, there has been significant growth of commercial ET in South America. Within North America, ET activity has been relatively constant in Holstein cattle, whereas there has been a large ET increase in the Angus breed and a concomitant ET decrease in some other beef breeds. Although a number of new technologies have been adopted within the ET industry in the last decade, the basic procedure of superovulation of donor cattle has undergone little improvement over the last 20 years. The export-import of frozen cattle embryos has become a well-established industry, governed by specific health regulations. The international movement of embryos is subject to sudden and dramatic disturbances, as exemplified by the 2001 outbreak of foot and mouth disease in Great Britain. It is probable that there will be an increased influence of animal rights issues on the ET industry in the future. Several companies in North America are currently commercially producing cloned cattle. The sexing of bovine semen with the use of flow cytometry is extremely accurate and moderate pregnancy rates in heifers have been achieved in field trials, but sexed semen currently is available in only a few countries and on an extremely limited basis. As of yet, all programs involving the production of transgenic cattle are experimental in nature.

Societal changes and the increasing desire and opportunity to preserve fertility have increased the demand for effective assisted reproductive technologies (ART) and have increased the range of scenarios in which ART is now used. In recent years, the "freeze-all" strategy of cryopreserving all oocytes or good quality embryos produced in an IVF cycle to transfer later-at a time that is more appropriate for reasons of medical need, efficacy, or desirability-has emerged as an accepted and valuable alternative to fresh embryo transfer. Indeed, improvements in cryopreservation techniques (vitrification) and the development of more efficient ovarian stimulation protocols have facilitated a dramatic increase in the practice of elective frozen embryo transfer (eFET). Alongside these advances, debate continues about whether eFET should be a standard treatment option available to the whole IVF population or if it is important to identify patient subgroups who are most likely to benefit from such an approach. Achieving successful outcomes in ART, whether by fresh or frozen embryo transfer, is influenced by a wide range of factors. As well as the efficiency of IVF and embryo transfer protocols and techniques, factors affecting implantation include maternal aging, sperm quality, the vaginal and endometrial microbiome, and peri-implantation levels of serum progesterone. The safety of eFET, both during ART cycles and on longer-term obstetric and neonatal outcomes, is also an important consideration. In this review, we explore the benefits and risks of freeze-all strategies in different scenarios. We review available evidence on the outcomes achieved with elective cryopreservation strategies and practices and how these compare with more traditional IVF cycles with fresh embryo transfers, both in the general IVF population and in subgroups of special interest. In addition, we consider how to optimize and individualize "freeze-all" procedures to achieve successful reproductive outcomes.Copyright © 2020 Bosch, De Vos and Humaidan.

After the first successful transfer of mammalian embryos in 1890, it was approximately 60 years before significant progress was reported in the basic technology of embryo transfer (ET) in cattle. Starting in the early 1970s, technology had progressed sufficiently to support the founding of commercial ET programs in several countries. Today, well-established and reliable techniques involving superovulation, embryo recovery and transfer, cryopreservation, and IVF are utilized worldwide in hundreds, if not thousands, of commercial businesses located in many countries. The mean number of embryos produced via superovulation has changed little in 40 years, but there have been improvements in synchrony and hormonal protocols. Cryopreservation of in vivo-derived embryos is a reliable procedure, but improvements are needed for biopsied and in vitro-derived embryos. High pregnancy rates are achieved when good quality embryos are transferred into suitable recipients and low pregnancy rates are often owing to problems in recipient management and not technology per se. In the future, unanticipated disease outbreaks and the ever-changing economics of cattle and milk prices will continue to influence the ET industry. The issue of abnormal pregnancies involving in vitro embryos has not been satisfactorily resolved and the involvement of abnormal epigenetics associate with this technology merits continued research. Last, genomic testing of bovine embryos is likely to be available in the foreseeable future. This may markedly decrease the number of embryos that are actually transferred and stimulate the evolution of more sophisticated ET businesses.Copyright © 2014 Elsevier Inc. All rights reserved.

【目的】探究添加胰岛素-转铁蛋白-亚硒酸钠(insulin-transferrin-sodium selenite,ITS)对牛体外胚胎生产、4 ℃低温保存及常规程序冷冻效果的影响,以期应用ITS提高牛体外胚胎的生产及保存效果。【方法】对屠宰场卵巢来源的牛卵母细胞进行体外受精,①研究在胚胎体外培养后期液M2中添加ITS对囊胚率的影响,试验分为M2中添加ITS组(M2+ITS)和M2中未添加ITS的对照组(M2-control);②研究在M2中添加ITS后进一步在前期培养液M1中添加ITS对早期胚胎发育的影响,试验分为M1和M2中均添加ITS组(M1+M2+ITS)和仅M2中添加ITS的对照组(M1-control);③在三气培养条件下对ITS改善胚胎发育的效果进行评估,试验分为添加ITS组(ITS)和未添加ITS的对照组(control),以上各组均统计卵裂率、7 d囊胚率及总囊胚率;④将获得的7 d牛囊胚在添加ITS的M2保存液(Hypo-ITS组)中低温保存(4 ℃)24 h后复苏培养,或者在添加ITS(Cryo-ITS组)的冷冻保存液中常规程序冷冻解冻后培养,以低温保存液中未添加ITS(Hypo-control组)和冷冻液中未添加ITS(Cryo-control组)为对照,统计胚胎的存活率和孵化率。【结果】与对照组相比,M2+ITS和M1+M2+ITS组的7 d囊胚率均显著提高(P＜0.05);三气培养条件下,ITS组7 d囊胚率显著提高(P＜0.05)。Hypo-ITS组4 ℃低温保存复苏的牛胚胎存活率与对照组差异不显著(P＞0.05),但孵化率显著高于对照组(P＜0.05);Cryo-ITS组的常规程序冷冻解冻的牛胚胎存活率及孵化率与对照组均无显著差异(P＞0.05)。【结论】ITS可作为添加剂及低温保护剂用于提高牛体外胚胎生产及低温保存效果,但在常规程序化冷冻液中添加ITS对牛体外胚胎常规程序冷冻效果的提升并不明显。

Cryopreservation of oocytes and embryos is a crucial step for widespread practical application of other techniques in domestic animal embryology. However, in contrast to the rapid development of procedures in the eighties and early nineties of the last century, new advancements with profound practical consequences have only been recently achieved. As a result of a long and controversial development, an alternative group of methods, vitrification, has proved its efficiency and practicality in the past few years. The aim of this short review is to characterize strategies to prevent cryoinjuries, summarize the development of vitrification, overview its recent achievements, and provide a perspective about possible application. Authors strongly believe that the future of mammalian oocyte and embryo cryopreservation will be mainly based on vitrification, and the rate of advancement will be determined by the rate by which embryologists learn and acknowledge this new approach.

During the past 50 years, the fertility of high-producing lactating dairy cows has decreased, associated with intensive selection for increased milk production. The physiological and metabolic changes associated with high milk production, including decreased (glucose, insulin, IGF-I) or increased (nonesterified fatty acids, ketone bodies) concentrations of circulating metabolites during nutrient partitioning associated with negative energy balance as well as uterine and nonuterine diseases have been linked with poor reproductive efficiency. Fertilization is typically above 80% and does not seem to be the principal factor responsible for the low fertility in dairy cows. However, early embryonic development is compromised in high-producing dairy cows, as observed by most embryonic losses occurring during the first 2 weeks after fertilization and may be linked to compromised oocyte quality due to a poor follicular microenvironment, suboptimal reproductive tract environment for the embryo, and/or inadequate maternal-embryonic communication. These and other factors related to embryo development will be discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

旨在探讨激素处理方案、供体牛月龄对高产荷斯坦奶牛超数排卵效果和性控体外胚胎生产效率的影响，为建立稳定高效且适用于高产荷斯坦奶牛的超数排卵方案以及提高高产奶牛性控胚胎体外生产效率提供技术支持。实验共选取高产荷斯坦奶牛60头，随机分为3组，分别按照注射FSH两周采卵一次(bi-weekly，BW)、注射FSH一周采卵一次(once weekly，OW)与不注射激素(control，CON)进行OPU-IVF体外性控胚胎生产，应用以上最佳方案进一步研究青年牛与经产牛在体外胚胎生产效率上是否存在差异。BW组的超排效果要优于OW组与CON组，BW组头均可用卵数(15.30±0.60)、可用卵占比(68.83%±1.92%)、头均A级卵数(9.86±0.38)与头均B级卵数(5.44±0.37)均显著高于OW组；BW组IVF早期胚胎体外发育优于OW组与CON组，其中BW组卵裂率(77.80%±1.08%)与头均胚胎数(5.11±0.27)均高于OW组与CON组且差异显著，BW组和CON组B级胚胎占比(13.95%±1.23%)均显著低于OW组；经产牛组头均卵子数(27.10±1.35)与头均胚胎数(5.29±0.28)均显著高于青年牛组，但卵子可用率经产牛组显著低于青年牛组，分别为64.44%±1.81%和75.01%±2.33%。BW组可获得最佳的超排效果，应用BW超排方案发现经产牛的体外胚胎生产效率优于青年牛。

体外胚胎冷冻保存技术是胚胎移植技术的重要组成部分，在辅助生殖技术中发挥重要作用，同时对种质资源保存、加强遗传改良和促进优质种源国际交流等方面具有重要意义。然而，体外胚胎冷冻过程中存在脂质含量过高、活性氧水平升高及机械损伤等问题，导致体外胚胎冷冻效率低，这极大地限制了体外胚胎冷冻保存技术的广泛应用。大量研究表明，通过去脂质、优化体外胚胎培养液、人工塌陷囊胚腔和优化冷冻程序等手段，可以有效提高冷冻后胚胎的存活率和发育能力。因此，本文概述了体外胚胎冷冻保存技术的研究进展和胚胎冷冻过程中存在的问题，总结了提高体外胚胎冷冻效率的方法措施，旨在为提高体外胚胎冷冻保存效率提供一定参考。