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Early ovine preantral follicles have a potential to grow until antral stage in two‐step culture system in the presence of aqueous extract of Justicia insularis
Author(s) -
Mbemya Gildas Tetaping,
Sá Naiza Arcângela Ribeiro,
Guerreiro Denise Damasceno,
Sousa Francisca Geovania Canafístula,
Nguedia Sylvain Njina,
Alves Benner Geraldo,
Santos Francielli Weber,
Pessoa Otília Deusdênia Loiola,
Comizzoli Pierre,
Figueiredo José Ricardo,
Rodrigues Ana Paula Ribeiro
Publication year - 2019
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.13477
Subject(s) - ovarian cortex , follicular phase , antral follicle , antrum , andrology , in vitro , biology , trehalose , in vitro maturation , oocyte , ovary , endocrinology , medicine , ovarian tissue , biochemistry , embryo , stomach , microbiology and biotechnology
Abstract The objective of this study was to determine whether preantral follicles cultured in vitro for 7 days within ovine ovarian cortical strips could be isolated at the secondary follicles (SF) and grown until antral stage during an additional 6 days period of in vitro culture in the presence of aqueous extract of Justicia insularis . Fresh ovarian fragments from 16 adult sheep were fixed for histological analysis (Control 1) or in vitro cultured individually in α‐MEM + supplemented with 0.3 mg/ml J. insularis (Step 1) for 7 days. Part of the fragments then were fixed for histological analysis (in vitro culture group). Remaining fragments were exposed stepwise to increasing trehalose concentrations before immediate isolation of SF and viability assessment (Control 2) or after 6 days of culture in α‐MEM ++ supplemented with 0.3 mg/ml J. insularis (Step 2). In Step 1, percentage of follicular activation was 80%. In Step 2, a significant increase ( p < 0.05) in follicular diameter and antrum formation within 6 days in vitro culture of isolated follicles was achieved. The total antioxidant capacity from both steps significantly increase ( p < 0.05) from day 2 to day 6. Confocal analysis of oocytes showed 57.14% oocytes with homogeneous distribution and 42.86% with peri‐cortical distribution. In conclusion, SF can be successfully isolated from sheep ovarian cortex after 7 days of culture and are capable of surviving and forming an antral cavity if cultured in vitro for an additional 6 days in the presence of 0.3 mg/ml J. insularis .