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Lower apoptosis rate in ovine preantral follicles from ovaries stored in supplemented preservation media

Published online by Cambridge University Press:  28 January 2015

R.J.S. Gonçalves ,
A.Y.P. Cavalcante ,
B.B. Gouveia ,
T.L.B. Lins ,
R.S. Barberino ,
V.G. Menezes ,
V.R.P. Barros ,
T.J.S. Macedo ,
J.R. Figueiredo  and
M.H.T. Matos
R.J.S. Gonçalves
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
A.Y.P. Cavalcante
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
B.B. Gouveia
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
T.L.B. Lins
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
R.S. Barberino
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
V.G. Menezes
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
V.R.P. Barros
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
T.J.S. Macedo
Affiliation:
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Laboratory of Cell Biology, Cytology and Histology, Federal University of San Francisco Valley (UNIVASF), Petrolina, Brazil.
J.R. Figueiredo
Affiliation:
Faculty of Veterinary Medicine, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceara, 60740-000, Fortaleza-CE, Brazil.
M.H.T. Matos*
Affiliation:
Universidade Federal do Vale do São Francisco (UNIVASF), Colegiado de Medicina Veterinária – Laboratório de Biologia Celular, Citologia e Histologia, Rodovia BR 407, Km 12, Lote 543 – Projeto de Irrigação Nilo Coelho – S/N, C1 CEP: 56300-990 – Petrolina – PE–Brasil.
*
All correspondence to: M.H.T. Matos. Universidade Federal do Vale do São Francisco (UNIVASF), Colegiado de Medicina Veterinária – Laboratório de Biologia Celular, Citologia e Histologia, Rodovia BR 407, Km 12, Lote 543 – Projeto de Irrigação Nilo Coelho – S/N, C1 CEP: 56300-990 – Petrolina – PE–Brasil. Tel.: +55 87 2101 4839. e-mail: helena.matos@univasf.edu.br
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Summary

The aim of this study was to investigate the effect of ovarian tissue transportation conditions (medium and period of time) on the morphology, apoptosis and development of ovine preantral follicles cultured in vitro. Each ovarian pair was cut into nine slices, with one fragment being fixed immediately (fresh control). The remaining fragments were placed individually in cryotubes containing conservation medium (minimal essential medium (MEM) without supplementation or MEM+ – with supplementation) and stored at 35ºC for 6 or 12 h without (non-cultured) or with subsequent culture for 5 days. Then, the fragments were processed for histological and terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labelling (TUNEL) examination. Preservation of ovarian slices in MEM or MEM+ (non-cultured) resulted in similar percentages of normal follicles when compared with the fresh control. Nevertheless, compared with the fresh control, a decrease in the percentage of normal follicles was observed in tissues cultured for 5 days. Only for tissues preserved in supplemented medium (MEM+) for 6 h, the percentage of TUNEL positive cells was similar between non-cultured tissues and tissues cultured for 5 days. Follicular activation and growth (follicular and oocyte diameter) were higher in cultured tissues than in fresh control or non-cultured tissues, except those from fragments preserved for 6 h in MEM and then cultured for 5 days in which no growth was observed. In conclusion, ovine ovarian tissue was successfully preserved in supplemented medium (MEM+) at a temperature close to physiological values (35°C) for up to 6 h without affecting apoptosis in the ovarian follicles and their ability to develop in vitro.

Keywords

GrowthOvaryPreservationSheepSurvival
Type
Research Article
Information
Zygote , Volume 23 , Issue 6 , December 2015 , pp. 943 - 950
Copyright
Copyright © Cambridge University Press 2015 

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