FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / In Vitro Cell Dev Biol Anim / In vitro culture supplementation of EGF for improving the su...
In vitro cellular & developmental biology. Animal2018; 54(10); 687-691; doi: 10.1007/s11626-018-0296-9

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

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.
Get Full Text
Publication Date: 2018-10-03 PubMed ID: 30284096DOI: 10.1007/s11626-018-0296-9Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research studies the impact of using various concentrations of epidermal growth factor (EGF) on the in vitro survival of horse ovarian follicles.

Research Background

  • The study explores folliculogenesis, a core process in the development and maturation of ovarian follicles, which is crucial for maintaining fertility.
  • Under natural conditions, 99.9% of an ovary’s follicles do not ovulate and undergo atresia, or natural death.
  • This research seeks to minimize this loss by understanding the mechanisms involved in the process and enhancing the in vitro follicular development by adding different concentrations of EGF, influencing its growth.

Research Methodology

  • The study carries out in vitro cultivation of equine (horse) preantral follicles, collected from a local slaughterhouse.
  • The team gathers 10 ovaries, treats them with 70% alcohol and PBS, then transports them for testing.
  • Each ovary is divided into 11 fragments of approximately 3 × 3 × 1mm, with one from each immediately fixed in Bouin (acting as the control group).
  • The remaining fragments go through individual cultivation for 2 and 6 days, with the additive of varying concentrations of EGF (0, 10, 50, 100, and 200 ng/mL).

Research Findings

  • The research finds a higher proportion of intact follicles with 0 and 100 ng/mL EGF concentrations after 2 days of culture. This means these EGF proportions promote better survival of ovarian follicles at this stage.
  • After 6 days of culture, only the 100 ng/mL EGF concentration displayed a significant difference when compared to the other EGF concentrations. The 100 ng/mL concentration proved to be the most effective in maintaining follicular survival out of all five concentrations.
  • Follicular development is observed after 2 days at all concentrations, proving that EGF fosters follicular survival in equines, most effectively at a concentration of 100 ng/mL in in vitro cultures. Additionally, this concentration is suggested for in-situ cultivations too.

Cite This Article

APA
Max MC, Bizarro-Silva C, Búfalo I, González SM, Lindquist AG, Gomes RG, Barreiros TRR, Lisboa LA, Morotti F, Seneda MM. (2018). In vitro culture supplementation of EGF for improving the survival of equine preantral follicles. In Vitro Cell Dev Biol Anim, 54(10), 687-691. https://doi.org/10.1007/s11626-018-0296-9

Publication

In vitro cellular & developmental biology. Animal
ISSN: 1543-706X
NlmUniqueID: 9418515
Country: Germany
Language: English
Volume: 54
Issue: 10
Pages: 687-691

Researcher Affiliations

Max, Marilu Constantino
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
Bizarro-Silva, Camila
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil. camilabizarros@gmail.com.
  • Laboratory Animal Reproduction, DCV, CCA, UEL, Londrina, PR, 86051-990, Cx. Postal: 10.011, Brazil. camilabizarros@gmail.com.
Búfalo, Isabela
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
González, Suellen Miguez
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
Lindquist, Andressa Guidugli
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
Gomes, Roberta Garbelini
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
Barreiros, Thales Ricardo Rigo
  • Department of Veterinary and Animal Production, Laboratory of Biotechnology of Animal Reproduction, State University of Northern Parana, Jacarezinho, Parana, Brazil.
Lisboa, Lívia Aires
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
Morotti, Fábio
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.
Seneda, Marcelo Marcondes
  • Department of Veterinary Clinical, Laboratory of Animal Reproduction, University of Londrina, Londrina, Parana, Brazil.

MeSH Terms

  • Animals
  • Cell Shape / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Culture Media / pharmacology
  • Epidermal Growth Factor / pharmacology
  • Female
  • Horses
  • Ovarian Follicle / cytology

References

This article includes 22 references
  1. Mao J, Smith MF, Rucker EB, Wu GM, McCauley TC, Cantley TC, Prather RS, Didion BA, Day BN. Effect of epidermal growth factor and insulin-like growth factor I on porcine preantral follicular growth, antrum formation, and stimulation of granulosal cell proliferation and suppression of apoptosis in vitro.. J Anim Sci 2004 Jul;82(7):1967-75.
    pubmed: 15309943doi: 10.2527/2004.8271967xgoogle scholar: lookup
  2. Celestino JJ, Bruno JB, Lima-Verde IB, Matos MH, Saraiva MV, Chaves RN, Martins FS, Lima LF, Name KP, Campello CC, Silva JR, Báo SN, Figueiredo JR. Recombinant epidermal growth factor maintains follicular ultrastructure and promotes the transition to primary follicles in caprine ovarian tissue cultured in vitro.. Reprod Sci 2009 Mar;16(3):239-46.
    pubmed: 19087984doi: 10.1177/1933719108325756google scholar: lookup
  3. Demeestere I, Centner J, Gervy C, Englert Y, Delbaere A. Impact of various endocrine and paracrine factors on in vitro culture of preantral follicles in rodents.. Reproduction 2005 Aug;130(2):147-56.
    pubmed: 16049152doi: 10.1530/rep.1.00648google scholar: lookup
  4. Haag KT, Magalhães-Padilha DM, Fonseca GR, Wischral A, Gastal MO, King SS, Jones KL, Figueiredo JR, Gastal EL. In vitro culture of equine preantral follicles obtained via the Biopsy Pick-Up method.. Theriogenology 2013 Apr 1;79(6):911-7.
    pubmed: 23434205doi: 10.1016/j.theriogenology.2013.01.001google scholar: lookup
  5. Andrade ER, Marcondes Seneda M, Alfieri AA, de Oliveira JA, Frederico Rodrigues Loureiro Bracarense AP, Figueiredo JR, Toniolli R. Interactions of indole acetic acid with EGF and FSH in the culture of ovine preantral follicles.. Theriogenology 2005 Sep 15;64(5):1104-13.
    pubmed: 16125554doi: 10.1016/j.theriogenology.2005.03.001google scholar: lookup
  6. Aguiar FLN, Lunardi FO, Lima LF, Bruno JB, Alves BG, Magalhães-Padilha DM, Cibin FWS, Berioni L, Apgar GA, Lo Turco EG, Gastal EL, Figueiredo JR. Role of EGF on in situ culture of equine preantral follicles and metabolomics profile.. Res Vet Sci 2017 Dec;115:155-164.
    pubmed: 28414979doi: 10.1016/j.rvsc.2017.04.001google scholar: lookup
  7. Gomes RG, Lisboa LA, Silva CB, Max MC, Marino PC, Oliveira RL, González SM, Barreiros TR, Marinho LS, Seneda MM. Improvement of development of equine preantral follicles after 6 days of in vitro culture with ascorbic acid supplementation.. Theriogenology 2015 Sep 15;84(5):750-5.
    pubmed: 26074067doi: 10.1016/j.theriogenology.2015.05.006google scholar: lookup
  8. Gonzalez SM, da Silva CB, Lindquist AG, Bufalo I, Morotti F, Lisboa LA, Seneda M. Regional distribution and integrity of equine ovarian pre-antral follicles.. Reprod Domest Anim 2017 Oct;52(5):836-841.
    pubmed: 28580754doi: 10.1111/rda.12986google scholar: lookup
  9. Pessoa AF, Rocha RM, Brito IR, Silva GM, Chaves RN, Magalhães-Padilha DM, Campello CC, Rodrigues AP, Nunes-Pinheiro DC, Figueiredo JR. Effect of morphological integrity, period, and type of culture system on the in vitro development of isolated caprine preantral follicles.. Theriogenology 2014 Jul 15;82(2):312-7.
    pubmed: 24839921doi: 10.1016/j.theriogenology.2014.04.008google scholar: lookup
  10. Erickson BH. Development and senescence of the postnatal bovine ovary.. J Anim Sci 1966 Aug;25(3):800-5.
    pubmed: 6007918doi: 10.2527/jas1966.253800xgoogle scholar: lookup
  11. Jimenez CR, Araújo VR, Penitente-Filho JM, de Azevedo JL, Silveira RG, Torres CA. The base medium affects ultrastructure and survival of bovine preantral follicles cultured in vitro.. Theriogenology 2016 Apr 1;85(6):1019-29.
    pubmed: 26711701doi: 10.1016/j.theriogenology.2015.11.007google scholar: lookup
  12. Araújo VR, Gastal MO, Figueiredo JR, Gastal EL. In vitro culture of bovine preantral follicles: a review.. Reprod Biol Endocrinol 2014 Aug 13;12:78.
    pubmed: 25117631doi: 10.1186/1477-7827-12-78google scholar: lookup
  13. Serafim MK, Duarte AB, Silva GM, Souza CE, Magalhães-Padilha DM, Moura AA, Silva LD, Campello CC, Figueiredo JR. Impact of growth hormone (GH) and follicle stimulating hormone (FSH) on in vitro canine preantral follicle development and estradiol production.. Growth Horm IGF Res 2015 Apr;25(2):85-9.
    pubmed: 25604894doi: 10.1016/j.ghir.2014.12.009google scholar: lookup
  14. Max MC, Silva CB, González SM, Lindquist AG, Búfalo I, Gomes RG, Morotti F, Costa CB, Barreiros T, Lisboa LA, Seneda MM. The development and integrity of equine pre-antral follicles cultured in vitro with follicle-stimulating hormone (FSH) supplementation.. Reprod Domest Anim 2017 Oct;52(5):899-904.
    pubmed: 28580717doi: 10.1111/rda.12996google scholar: lookup
  15. Zhou H, Zhang Y. Effect of growth factors on in vitro development of caprine preantral follicle oocytes.. Anim Reprod Sci 2005 Dec;90(3-4):265-72.
    pubmed: 16298274doi: 10.1016/j.anireprosci.2005.01.008google scholar: lookup
  16. Duarte AB, Araújo VR, Chaves RN, da Silva GM, Luz VB, Haag KT, Magalhães-Padilha DM, Almeida AP, Lobo CH, Campello CC, de Figueiredo JR. Insulin-like growth factor II (IGF-II) and follicle stimulating hormone (FSH) combinations can improve the in vitro development of grown oocytes enclosed in caprine preantral follicles.. Growth Horm IGF Res 2013 Feb-Apr;23(1-2):37-44.
    pubmed: 23333247doi: 10.1016/j.ghir.2012.12.002google scholar: lookup
  17. Markström E, Svensson ECh, Shao R, Svanberg B, Billig H. Survival factors regulating ovarian apoptosis -- dependence on follicle differentiation.. Reproduction 2002 Jan;123(1):23-30.
    pubmed: 11869183doi: 10.1530/rep.0.1230023google scholar: lookup
  18. Driancourt MA, Paris A, Roux C, Mariana JC, Palmer E. Ovarian follicular populations in pony and saddle-type mares.. Reprod Nutr Dev (1980) 1982;22(6):1035-47.
    pubmed: 7163613doi: 10.1051/rnd:19820714google scholar: lookup
  19. Celestino JJ, Bruno JB, Saraiva MV, Rocha RM, Brito IR, Duarte AB, Araújo VR, Silva CM, Matos MH, Campello CC, Silva JR, Figueiredo JR. Steady-state level of epidermal growth factor (EGF) mRNA and effect of EGF on in vitro culture of caprine preantral follicles.. Cell Tissue Res 2011 Jun;344(3):539-50.
    pubmed: 21503599doi: 10.1007/s00441-011-1162-1google scholar: lookup
  20. Saraiva MV, Rossetto R, Brito IR, Celestino JJ, Silva CM, Faustino LR, Almeida AP, Bruno JB, Magalhães DM, Matos MH, Campello CC, Figueiredo JR. Dynamic medium produces caprine embryo from preantral follicles grown in vitro.. Reprod Sci 2010 Dec;17(12):1135-43.
    pubmed: 20926838doi: 10.1177/1933719110379269google scholar: lookup
  21. Gutierrez CG, Ralph JH, Telfer EE, Wilmut I, Webb R. Growth and antrum formation of bovine preantral follicles in long-term culture in vitro.. Biol Reprod 2000 May;62(5):1322-8.
    pubmed: 10775183doi: 10.1095/biolreprod62.5.1322google scholar: lookup
  22. Silva JR, van den Hurk R, Costa SH, Andrade ER, Nunes AP, Ferreira FV, Lôbo RN, Figueiredo JR. Survival and growth of goat primordial follicles after in vitro culture of ovarian cortical slices in media containing coconut water.. Anim Reprod Sci 2004 Apr;81(3-4):273-86.
    pubmed: 14998653doi: 10.1016/j.anireprosci.2003.09.006google scholar: lookup

Citations

This article has been cited 4 times.
  1. Ahmed AE, Sindi RA, Yousef NA, Hussein HA, Badr MR, Syaad KMA, Al-Saeed FA, Hassaneen ASA, Abdelrahman M, Ali ME. Impact of epidermal growth factor and/or β-mercaptoethanol supplementations on the in vitro produced buffaloes' embryos.. Front Vet Sci 2023;10:1138220.
    doi: 10.3389/fvets.2023.1138220pubmed: 36992972google scholar: lookup
  2. Taghizabet N, Bahmanpour S, Zarei-Fard N, Mohseni G, Aliakbari F, Dehghani F. Effect of endometrial cell-conditioned medium and platelet-rich plasma on the developmental competence of mouse preantral follicles: An in vitro study.. Clin Exp Reprod Med 2022 Sep;49(3):175-184.
    doi: 10.5653/cerm.2022.05260pubmed: 36097733google scholar: lookup
  3. Mo J, Sun L, Cheng J, Lu Y, Wei Y, Qin G, Liang J, Lan G. Non-targeted Metabolomics Reveals Metabolic Characteristics of Porcine Atretic Follicles.. Front Vet Sci 2021;8:679947.
    doi: 10.3389/fvets.2021.679947pubmed: 34381832google scholar: lookup
  4. Simon LE, Kumar TR, Duncan FE. In vitro ovarian follicle growth: a comprehensive analysis of key protocol variables†.. Biol Reprod 2020 Aug 21;103(3):455-470.
    doi: 10.1093/biolre/ioaa073pubmed: 32406908google scholar: lookup
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.