Development of sheep secondary follicles and preservation of aromatase and metalloproteinases 2 and 9 after vitrification and in vitro culture

Francisco Denilson Rodrigues Gomes, Danielle Cristina Calado de Brito, Naíza Arcângela Ribeiro de Sá, Lucy Vanessa Sulca Ñaupas, Gaby Judith Quispe Palomino, Renato Felix da Silva, Éverton Pimentel Ferreira Lopes, Gildas Tetaping Mbemya, Benner Geraldo Alves, Mary Zelinski, José Ricardo de Figueiredo, Ana Paula Ribeiro Rodrigues

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


The cryopreservation of secondary follicles (SF) is a promising alternative to preserve the reproductive potential both in humans and animals in situations in which the transplantation of ovarian tissue is not possible. The objective of the present study was cryopreserved SF isolated sheep. Beyond follicular morphology, viability and development, we investigated proteins related to steroidogenic function and basement membrane remodeling [metalloproteinases 2 (MMP-2) and 9 (MMP-9)] in fresh SF (FSF) and vitrified SF (VSF) followed by in vitro culture for 6 (D6) or 12 days (D12). The percentage of intact follicles, follicular and oocyte diameter of the VSF were lower than FSF on both days of culture (P < 0.05). The VSF viability was statistically reduced from D6 (95.5%) to D12 (77.3%) but did not differ from the FSF on both days (D6:96.2% to D12:86.5%). Antrum formation in the VSF (D6: 59.13%; D12: 79.56%) was significantly lower than the FSF (D6: 79.61%; D12: 92.23%). However, an increase in this percentage was observed from D6 to D12 in both groups. Aromatase showed stronger labeling on FSF D6 and VSF D12 compared to other treatments (P < 0.05). MMP-2 showed a similar pattern of labeling in FSF D6 and VSF D12, similarly to that observed in FSF D12 and VSF D6. MMP-9 was similar in FSF and VSF cultivated for 6 and 12 days. In conclusion, VSF are able to grow and develop during 12 days of in vitro culture and showed evidence of preservation of steroidogenic function and remodeling of the basement membrane.

Original languageEnglish (US)
Pages (from-to)247-259
Number of pages13
JournalCell and Tissue Banking
Issue number2
StatePublished - Jun 2022


  • Aromatase
  • Fertility
  • Metalloproteinase
  • Preantral follicle
  • Vitrification

ASJC Scopus subject areas

  • Biomaterials
  • Biomedical Engineering
  • Cell Biology
  • Transplantation


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