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 |
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Authors: | Gildas Tetaping Mbemya,Naiza Arc ngela Ribeiro de S ,Denise Damasceno Guerreiro,Francisca Geovania Canafí stula de Sousa,Sylvain Njina Nguedia,Benner Geraldo Alves,Francielli Weber Santos,Otí lia Deusdê nia Loiola Pessoa,Pierre Comizzoli,Jos Ricardo Figueiredo,Ana Paula Ribeiro Rodrigues |
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Affiliation: | Gildas Tetaping Mbemya,Naiza Arcângela Ribeiro de Sá,Denise Damasceno Guerreiro,Francisca Geovania Canafístula de Sousa,Sylvain Njina Nguedia,Benner Geraldo Alves,Francielli Weber Santos,Otília Deusdênia Loiola Pessoa,Pierre Comizzoli,José Ricardo Figueiredo,Ana Paula Ribeiro Rodrigues |
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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. |
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Keywords: | antral follicles early secondary follicles in vitro culture
Justicia insularis
ovarian cortex |
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