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. 2021 May;7(3):868-875.
doi: 10.1002/vms3.428. Epub 2021 Jan 27.

Evaluation of a small volume oil-free in vitro production system for bovine embryos

Carina Blaschka  1 , Sophie Diers  1 , Mariya Aravina  1 , Swantje Geisler  1 , Gerhard Schuler  2 , Jens Tetens  1
Affiliations

Evaluation of a small volume oil-free in vitro production system for bovine embryos

Carina Blaschka et al. Vet Med Sci. 2021 May.

Abstract

Background: Embryos are usually produced in culture systems with an oil overlay, which conveys protection against the evaporation of water and microbial contamination. The oil can also release toxic substances and absorb essential components, such as hormones, which adversely affect the quality of the oocytes and the development of embryos in vitro.

Objective: The aim of this study was to validate an oil-free bovine in vitro production (IVP) system.

Method: Cumulus-oocyte complexes collected from abattoir-derived ovaries were matured, fertilized and cultured employing a standard system. The quantity of medium in both groups (with and without an oil overlay) and throughout all stages of IVP was maintained at a volume of 100 μl. The oil group was covered with paraffin oil. The maturation stage of oocytes was assessed using fluorescence staining after 24 hr and developmental stages of embryos were evaluated on day 8. The expanded day 8 blastocysts were assessed by live-dead staining.

Results: Oocytes matured in the absence of an oil overlay had significantly higher maturation rates when compared against matured oocytes in medium with an oil overlay. Steroid concentration is higher in medium after maturation without oil cover. The developmental rate was significantly higher after culture without oil overlay. The total cell number and the live-dead ratio was not significantly different. The osmolality did not differ between both groups during maturation and slightly decreased during culture without oil.

Conclusion: Based on the current study, bovine oil-free IVP systems can be suggested as an alternative to oil-covered medium.

Keywords: bovine; in vitro production; oil overlay.

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Conflict of interest statement

We confirm that the manuscript has been read and approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed. We further confirm that the order of authors listed in the manuscript has been approved by all authors.

Figures

FIGURE 1
FIGURE 1
Maturation rate of COCs after 24 hr and development rate on day 8 with or without an oil overlay in the entire IVP (mean ± SD; a:b p ≤ 0.05)
FIGURE 2
FIGURE 2
Steroid concentration in IVM medium after 24 hr of maturation (E2: 17ß‐oestradiol, p = 0.099; P4: progesterone, a:b p ≤ 0.05; mean ± SD)
FIGURE 3
FIGURE 3
Total cell number and live–dead ratio of expanded blastocysts after IVP with or without oil respectively (with oil: n = 25; without oil: n = 26; mean ± SD; p > 0.05)
See this image and copyright information in PMC

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