TY - JOUR
T1 - Vitrification of caprine secondary and early antral follicles as a perspective to preserve fertility function
AU - Lopes, Everton Pimentel Ferreira
AU - Rodrigues, Giovanna Quintino
AU - de Brito, Danielle Cristina Calado
AU - Rocha, Rebeca Magalhães Pedrosa
AU - Ferreira, Anna Clara Accioly
AU - de Sá, Naíza Arcângela Ribeiro
AU - Silva, Renato Félix da
AU - de Alcântara, Gabriel Las Heras
AU - Alves, Benner Geraldo
AU - Figueiredo, José Ricardo de
AU - Zelinski, Mary
AU - Rodrigues, Ana Paula Ribeiro
N1 - Funding Information:
Funding for this work was provided by National Council of Technological and Scientific Development (CNPq: 437458/2018-0 ). Everton Pimentel Ferreira Lopes is a recipient of a grant from CNPq. Ana Paula Ribeiro Rodrigues is the recipient of a grant (Number of the process: 308.071/2016-6) from CNPq. This work was also supported by a Fulbright Visiting Professor Scholarship to Ana Paula Ribeiro Rodrigues, NIH R01HD083930 to M.B. Zelinski and NIH P51OD011092 to the Oregon National Primate Research Center.
Publisher Copyright:
© 2020 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn
PY - 2020/9
Y1 - 2020/9
N2 - The present study aimed to evaluate the structure, survival and development of isolated caprine (secondary-SEC and early antral-EANT) follicles, after vitrification in the presence of synthetic polymers and in vitro culture. Additionally, transzonal projections (TZPs) and p450 aromatase enzyme were evaluated. After isolation, SEC and EANT follicles were in vitro cultured for six days or vitrified. After one week, SEC and EANT follicles were warmed and also in vitro cultured for six days. Data revealed that the percentage of morphologically normal follicles was similar between fresh and vitrified follicles in both follicular categories and antrum formation rate was similar between fresh and vitrified SEC follicles. Fluorescence by calcein-AM did not show difference between fresh and vitrified (SEC and EANT) follicles, however, the trypan blue test showed low viability for vitrified follicles. The integrity of TZPs was not affected between fresh and vitrified SEC follicles, however, in vitrified EANT follicles, there were signs of TZPs loss. Regarding steroidogenic function, it was observed a positive staining for p450 aromatase enzyme in fresh and vitrified SEC and EANT follicles. It was concluded that SEC follicles seem to be more resistant to vitrification than EANT follicles, as shown by the trypan blue test and TZPs assay. Future studies may confirm this hypothesis, in order to consolidate the use of SEC and EANT follicles as an alternative to ovary cryopreservation.
AB - The present study aimed to evaluate the structure, survival and development of isolated caprine (secondary-SEC and early antral-EANT) follicles, after vitrification in the presence of synthetic polymers and in vitro culture. Additionally, transzonal projections (TZPs) and p450 aromatase enzyme were evaluated. After isolation, SEC and EANT follicles were in vitro cultured for six days or vitrified. After one week, SEC and EANT follicles were warmed and also in vitro cultured for six days. Data revealed that the percentage of morphologically normal follicles was similar between fresh and vitrified follicles in both follicular categories and antrum formation rate was similar between fresh and vitrified SEC follicles. Fluorescence by calcein-AM did not show difference between fresh and vitrified (SEC and EANT) follicles, however, the trypan blue test showed low viability for vitrified follicles. The integrity of TZPs was not affected between fresh and vitrified SEC follicles, however, in vitrified EANT follicles, there were signs of TZPs loss. Regarding steroidogenic function, it was observed a positive staining for p450 aromatase enzyme in fresh and vitrified SEC and EANT follicles. It was concluded that SEC follicles seem to be more resistant to vitrification than EANT follicles, as shown by the trypan blue test and TZPs assay. Future studies may confirm this hypothesis, in order to consolidate the use of SEC and EANT follicles as an alternative to ovary cryopreservation.
KW - Cryopreservation
KW - Endocrine function
KW - Goat
KW - Ovarian follicles
KW - TZPs
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U2 - 10.1016/j.repbio.2020.05.001
DO - 10.1016/j.repbio.2020.05.001
M3 - Article
C2 - 32418820
AN - SCOPUS:85084554792
SN - 1642-431X
VL - 20
SP - 371
EP - 378
JO - Reproductive Biology
JF - Reproductive Biology
IS - 3
ER -