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In vitro culture supplementation of EGF for improving the survival of equine preantral follicles

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Abstract

Folliculogenesis is a process of development and maturation of the ovarian follicles, being essential for the maintenance of fertility. In in vivo conditions, 99.9% of the follicles of an ovary do not ovulate and undergo atresia. In order to minimize this loss and to clarify the existing mechanisms, a technique was developed that allows for the in vitro follicular development. The objective of this study was to evaluate the effects of different epidermal growth factor (EGF) concentrations on the in vitro culturing of equine preantral follicles. Ovaries (n = 10) were collected from a local slaughterhouse of mares in seasonal anestrus, washed with 70% alcohol and PBS, and transported. The inner portion of the ovary was divided into 11 fragments of approximately 3 × 3 × 1 mm. A fragment of each ovary was immediately fixed in Bouin (control group). The remaining 10 fragments were individually cultured for 2 and 6 d. The medium was supplemented with different concentrations of EGF (0, 10, 50, 100, and 200 ng/mL). After cultivation, the fragments were processed and classified according to the developmental stage and morphology. In total, 1065 slides containing 6105 tissue sections were evaluated. Within 2 d of culture, there was a higher proportion of intact follicles at the EGF concentrations of 0 and 100 ng/mL (p > 0.05). After 6 d of culture, only the EGF concentration of 100 ng/mL demonstrated a difference when compared to the other treatments (0, 10, 50 and 200 ng/mL of EGF, p > 0.05). There was follicular development after 2 d at all EGF concentrations. Thus, we suggest that EGF promotes follicular survival in equines at a concentration of 100 ng/mL in in vitro cultures of ovarian fragments for 2 d. In addition, we suggest that EGF promotes follicular survival in equines at a concentration of 100 ng/mL in situ cultivation.

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Author notes

  1. Camila Bizarro-Silva

    Present address: Laboratory Animal Reproduction, DCV, CCA, UEL, Londrina, PR, 86051-990, Cx. Postal: 10.011, Brazil

Authors and Affiliations

  1. Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil

    Marilu Constantino Max, Camila Bizarro-Silva, Isabela Búfalo, Suellen Miguez González, Andressa Guidugli Lindquist, Roberta Garbelini Gomes, Lívia Aires Lisboa, Fábio Morotti & Marcelo Marcondes Seneda

  2. Department of Veterinary and Animal Production, Laboratory of Biotechnology of Animal Reproduction, State University of Northern Parana, Jacarezinho, Parana, Brazil

    Thales Ricardo Rigo Barreiros

Authors
  1. Marilu Constantino Max
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  2. Camila Bizarro-Silva
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  3. Isabela Búfalo
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  4. Suellen Miguez González
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  5. Andressa Guidugli Lindquist
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  6. Roberta Garbelini Gomes
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  7. Thales Ricardo Rigo Barreiros
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  8. Lívia Aires Lisboa
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  9. Fábio Morotti
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  10. Marcelo Marcondes Seneda
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Correspondence to Camila Bizarro-Silva.

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Editor: Tetsuji Okamoto

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Max, M.C., Bizarro-Silva, C., Búfalo, I. et al. In vitro culture supplementation of EGF for improving the survival of equine preantral follicles. In Vitro Cell.Dev.Biol.-Animal 54, 687–691 (2018). https://doi.org/10.1007/s11626-018-0296-9

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