Abstract
Conventionally, in vitro–fertilized (IVF) bovine embryos are morphologically evaluated at the time of embryo transfer to select those that are likely to establish a pregnancy. This method is, however, subjective and results in unreliable selection. Here we describe a novel selection system for IVF bovine blastocysts for transfer that traces the development of individual embryos with time-lapse cinematography in our developed microwell culture dish and analyzes embryonic metabolism. The system can noninvasively identify prognostic factors that reflect not only blastocyst qualities detected with histological, cytogenetic, and molecular analysis but also viability after transfer. By assessing a combination of identified prognostic factors—(i) timing of the first cleavage; (ii) number of blastomeres at the end of the first cleavage; (iii) presence or absence of multiple fragments at the end of the first cleavage; (iv) number of blastomeres at the onset of lag-phase, which results in temporary developmental arrest during the fourth or fifth cell cycle; and (v) oxygen consumption at the blastocyst stage—pregnancy success could be accurately predicted (78.9%). The conventional method or individual prognostic factors could not accurately predict pregnancy. No newborn calves showed neonatal overgrowth or death. Our results demonstrate that these five predictors and our system could provide objective and reliable selection of healthy IVF bovine embryos.
Highlights
The criteria for selecting bovine in vitro fertilized (IVF) embryos for transfer are based on morphological quality at the time of transfer [1]
Hypoxic conditions are suitable for microwell culture dish, and we used 5% O2 in subsequent studies
The lag-phase was observed in 99.6% of embryos that reached the blastocyst stage; the number of blastomeres at the onset of the phase varied from 4 to 16, and embryos were classified into three groups: 4–5, 6–8, and 9–16 blastomeres (Fig. S1)
Summary
The criteria for selecting bovine in vitro fertilized (IVF) embryos for transfer are based on morphological quality at the time of transfer [1]. This approach is widely considered extremely subjective and inadequate [2]. Novel criteria allowing objective and reliable selection of embryos for transfer are required to advance bovine IVF technology. Noninvasive criteria that could predict blastocyst qualities and viability may lead to novel methods for selecting bovine embryos for transfer
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