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Vitrified sheep isolated secondary follicles are able to grow and form antrum after a short period of in vitro culture

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Abstract

The risk of reintroducing malignant cells after ovarian graft into patients following post-cancer treatment is an obstacle for clinical applications (autotransplantation). In this context, in vitro follicle culture would be an alternative to transplantation in order to minimize such risks. Therefore, the aim of this study was to compare the development of secondary follicles after vitrification in isolated form (without stroma) with vitrification in in situ form (within fragments of ovarian tissue). Follicles were first isolated from ovarian fragments from mixed-breed ewes and then vitrified; these comprised the Follicle-Vitrification group (Follicle-Vit), or fragments of ovarian tissue were first vitrified, followed by isolation of the follicles, resulting in the Tissue-Vitrification group (Tissue-Vit). Control and vitrified groups were submitted to in vitro culture (6 days) and follicular morphology, viability, antrum formation, follicle and oocyte diameter, growth rate, ultrastructural characteristics and cell proliferation were evaluated. The percentages of morphologically normal follicles and antrum formation were similar among groups. Follicular viability and oocyte diameter were similar between Follicle-Vit and Tissue-Vit. The follicular diameter and growth rate of Follicle-Vit were similar to the Control, while those of Tissue-Vit were significantly lower compared to the Control. Both vitrified groups had an augmented rate of granulosa cellular proliferation compared to Control. Secondary follicles can be successfully vitrified before or after isolation from the ovarian tissue without impairing their ability to survive and grow during in vitro culture.

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Financial support

This work was carried out at the Laboratory of Manipulation of Oocytes and Ovarian Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine of Ceará State University, Fortaleza, CE, Brazil, and supported by CNPq. Franciele Osmarini Lunardi is a recipient of a grant from CAPES Brazil. In addition, Ana Paula Ribeiro Rodrigues and José Ricardo de Figueiredo are recipients of a grant from CNPq Brazil.

Conflict of interest

The authors declare that there is no potential conflict of interest.

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Authors and Affiliations

  1. Laboratory of Manipulation of Oocytes and Ovarian Pre-antral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, Ceará State University, Fortaleza, CE, Brazil

    Franciele Osmarini Lunardi, Laritza Ferreira de Lima, Valdevane Rocha Araújo, Ivina Rocha Brito, Cláudio Cabral Campello, José Ricardo de Figueiredo & Ana Paula Ribeiro Rodrigues

  2. Health Center, University of Fortaleza (UNIFOR), Edson Queiroz, Fortaleza, Ceará, Brazil

    Roberta Nogueira Chaves

  3. State University of Ceara, Fortaleza, Ceara, Brazil

    Carlos Eduardo Azevedo Souza

  4. Laboratório de Ultraestrutura, CPqAM-FIOCRUZ, Universidade Federal de Pernambuco, Recife, PE, Brazil

    Mariana Aragão Matos Donato & Christina Alves Peixoto

  5. Molecular Animal Biotechnology Laboratory, Szent Istvan University, Gödöllö, Hungary

    Andras Dinnyes

Authors
  1. Franciele Osmarini Lunardi
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  2. Roberta Nogueira Chaves
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  3. Laritza Ferreira de Lima
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  4. Valdevane Rocha Araújo
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  5. Ivina Rocha Brito
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  6. Carlos Eduardo Azevedo Souza
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  7. Mariana Aragão Matos Donato
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  8. Christina Alves Peixoto
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  9. Andras Dinnyes
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  10. Cláudio Cabral Campello
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  11. José Ricardo de Figueiredo
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  12. Ana Paula Ribeiro Rodrigues
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Correspondence to Franciele Osmarini Lunardi.

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Lunardi, F.O., Chaves, R.N., de Lima, L.F. et al. Vitrified sheep isolated secondary follicles are able to grow and form antrum after a short period of in vitro culture. Cell Tissue Res 362, 241–251 (2015). https://doi.org/10.1007/s00441-015-2181-0

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  • DOI: https://doi.org/10.1007/s00441-015-2181-0

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