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RESEARCH ARTICLE

High insulin concentrations promote the in vitro growth and viability of canine preantral follicles

Michelle K. B. Serafim A D , Gerlane M. Silva A , Ana B. G. Duarte A , V. R. Araújo A , T. F. P. Silva A , A. K. F. Lima C , R. N. Chaves A , C. C. Campello A , L. D. M. Silva B and J. R. Figueiredo A
+ Author Affiliations
- Author Affiliations

A Faculty of Veterinary Medicine, Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Veterinary Science Post Graduation Program (VSPGP), State University of Ceara, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60.740-000, Brazil.

B Faculty of Veterinary Medicine, Laboratory of Carnivore Reproduction (LRC), State University of Ceara, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, CE 60.740-000, Brazil.

C Faculty of Veterinary Medicine, Federal University of Tocantins, Araguaina, TO, Brazil.

D Corresponding author. Email: michellekarenbs@hotmail.com

Reproduction, Fertility and Development 25(6) 927-934 https://doi.org/10.1071/RD12074
Submitted: 11 March 2012  Accepted: 4 August 2012   Published: 31 August 2012

Abstract

To determine whether the effects of different concentrations of insulin on the development of canine preantral follicles in vitro were associated or not with FSH, secondary follicles were isolated and cultured. In Experiment 1, follicles were cultured in the following media: modified minimum essential medium (CtrlMEM) alone; CtrlMEM plus 5 ng mL–1 insulin (Ins5ng); CtrlMEM plus 10 ng mL–1 insulin (Ins10ng); and CtrlMEM plus 10 μg mL–1 insulin. In Experiment 2, follicles were cultured in the same media but in the presence of sequential FSH (i.e. CtrlFSH, Ins5ngF, Ins10ngF and 10μgF, respectively). Increasing concentrations of FSH (100, 500 and 1000 ng mL–1) were added sequentially to the culture medium on Days 0, 6 and 12 of culture. Viability were assessed at the end of culture and follicular diameter and the antrum formation rate at four time points (Days 0, 6, 12 and 18). In Experiment 1, the high insulin concentration significantly increased follicular viability (P < 0.05). In contrast, in Experiment 2, viability was not affected by the inclusion of insulin. In addition, viability was significantly better in follicles cultured in CtrlFSH (P < 0.05). The diameter of follicles in the high-insulin group in Experiment 1 and high-insulin plus FSH group in Experiment 2 was superior to other groups tested. In experiment 2, the Ins10μg and Ins10μgF groups exhibited significantly higher antrum formation rates than the other groups. In conclusion, in the absence of FSH, high concentrations of insulin have beneficial effects on follicular viability. However, to promote the growth of canine preantral follicles in vitro, it is recommended that a combination of insulin and FSH be added to the medium.

Additional keyword: FSH.


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