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Effects of vitrification on the viability of alginate encapsulated isolated bovine pre-antral follicles

  • Fertility Preservation
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Individual follicle cryopreservation techniques, without hydrogel support, are labor-intensive and a substantial proportion of isolated follicles are lost during handling and after warming. Therefore, the viability and morphology of isolated bovine (as a model for human) pre-antral follicles after vitrification and warming, when encapsulated in alginate beads, were investigated.

Methods

Bovine pre-antral follicles were mechanically isolated and divided into four different groups: (1) culture in 2% alginate beads (3D system) and vitrification in beads using mesh cups (3DVIT), (2) culture in 2% alginate beads (3DCUL), (3) culture in 96-well plates (2D system) and vitrification using High Security Vitrification straws® (2DVIT), (4) culture in a 2D system (2DCUL). The same vitrification and warming protocols were used for embedded (3DVIT) and non-embedded follicles (2DVIT).

Results

No differences were observed in follicle viability between group 2DCUL and 3DCUL. Group 3DVIT showed the lowest viability (45.9%) according to calcein and neutral red staining among all groups. Group 2DVIT displayed the highest viability (87.5%) and largest percentage of follicles with a well-preserved morphology.

Conclusions

Our results show that, using a vitification protocol optimized for non-embedded follicles, 2D culture is more effective in vitrifying isolated follicles. However, embedding in alginate allow to handle follicles more efficiently, i.e., without excessive manipulation and thus less labor-intensive in combination with a reduced loss of follicles during the procedure. Based on the increased work efficiency, but lower viability and higher proportion of follicles showing impaired morphology, we consider it advantageous to optimize the protocol for the vitrification of embedded follicles to increase survival and maintain morphology after vitrification.

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Acknowledgements

The authors thank Silke Andries, Els Merckx and Katty Huybrechts for their excellent technical assistance and the local slaughterhouses for their cooperation in sample collection.

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All (co-)authors state that the funding of this research is provided by the independent Operational Costs of the University of Antwerp.

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Authors and Affiliations

  1. Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, Laboratory of Veterinary Physiology and Biochemistry, Gamete Research Centre, University of Antwerp, Universiteitsplein 1, Gebouw U, 2610, Wilrijk, Belgium

    Anniek Bus, Veerle van Hoeck, An Langbeen, Jo L. M. R. Leroy & Peter E. J. Bols

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  1. Anniek Bus
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  2. Veerle van Hoeck
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  3. An Langbeen
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Correspondence to Anniek Bus.

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Bus, A., van Hoeck, V., Langbeen, A. et al. Effects of vitrification on the viability of alginate encapsulated isolated bovine pre-antral follicles. J Assist Reprod Genet 35, 1187–1199 (2018). https://doi.org/10.1007/s10815-018-1208-3

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