TY - JOUR
T1 - Development of sheep secondary follicles and preservation of aromatase and metalloproteinases 2 and 9 after vitrification and in vitro culture
AU - Gomes, Francisco Denilson Rodrigues
AU - de Brito, Danielle Cristina Calado
AU - de Sá, Naíza Arcângela Ribeiro
AU - Ñaupas, Lucy Vanessa Sulca
AU - Palomino, Gaby Judith Quispe
AU - da Silva, Renato Felix
AU - Lopes, Éverton Pimentel Ferreira
AU - Mbemya, Gildas Tetaping
AU - Alves, Benner Geraldo
AU - Zelinski, Mary
AU - de Figueiredo, José Ricardo
AU - Rodrigues, Ana Paula Ribeiro
N1 - Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer Nature B.V.
PY - 2022/6
Y1 - 2022/6
N2 - 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.
AB - 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.
KW - Aromatase
KW - Fertility
KW - Metalloproteinase
KW - Preantral follicle
KW - Vitrification
UR - http://www.scopus.com/inward/record.url?scp=85130739232&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85130739232&partnerID=8YFLogxK
U2 - 10.1007/s10561-021-09937-5
DO - 10.1007/s10561-021-09937-5
M3 - Article
C2 - 34152507
AN - SCOPUS:85130739232
SN - 1389-9333
VL - 23
SP - 247
EP - 259
JO - Cell and Tissue Banking
JF - Cell and Tissue Banking
IS - 2
ER -