In vitro culture of equine preantral follicles obtained via the Biopsy Pick-Up method.
Abstract: The objective was to conduct a preliminary evaluation of the efficacy of two media for in vitro culture of equine preantral follicles. Ovarian cortical strips were obtained from mares (N = 10) via the Biopsy Pick-Up method during the breeding season. Ovarian tissue was immediately submitted to histological analysis (noncultured control; D0) or cultured in situ for 1 day (D1) or 7 days (D7) in either α-MEM or TCM-199 and submitted to histological analysis, generating five treatment groups: noncultured control, α-MEM:D1, TCM-199:D1, α-MEM:D7, and TCM-199:D7. Preantral follicles were evaluated for follicle class (primordial, transitional, primary, and secondary) and morphology (normal vs. abnormal). A total of 142 preantral follicles were analyzed in five replicates. No follicles were observed in the TCM-199:D7 treatment group. The proportion of primordial follicles was higher (P < 0.03) in the control compared to the α-MEM:D7 treatment group. The proportion of primary follicles was higher (P < 0.04) in the α-MEM:D7 treatment group compared to the control. The proportion of developing follicles (transitional, primary, and secondary) was higher (P < 0.03) in the α-MEM:D7 treatment group compared to the control group. There was a greater (P < 0.004) percentage of morphologically normal developing follicles in the α-MEM:D1 treatment group compared to the TCM-199:D1 treatment group. Overall, the percentage of morphologically normal follicles was higher in the control group (72%; P < 0.02) and α-MEM:D1 group (84%; P < 0.0001) compared to the α-MEM:D7 (27%) treatment group. Mean follicle diameter was greater (P < 0.04) in the α-MEM:D7 treatment group (40.6 ± 1.1 μm) compared to the control group (37.3 ± 0.7 μm). Mean oocyte diameter was greater in the α-MEM:D1 (31.0 ± 0.7 μm; P < 0.006), TCM-199:D1 (30.7 ± 1.8 μm; P < 0.006), and α-MEM:D7 (33.2 ± 1.8 μm; P < 0.006) treatment groups compared to the control group (27.4 ± 0.9 μm). In conclusion, based on these preliminary data, in vitro culture of equine ovarian fragments obtained in vivo via the Biopsy Pick-Up method promoted preantral follicle development and follicle and oocyte growth in α-MEM for 7 days, with some follicles remaining morphologically normal throughout the culture period.
Copyright © 2013 Elsevier Inc. All rights reserved.
Publication Date: 2013-02-20 PubMed ID: 23434205DOI: 10.1016/j.theriogenology.2013.01.001Google Scholar: Lookup
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- Comparative Study
- Evaluation Study
- Journal Article
Summary
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The research study investigates the effectiveness of two different media, α-MEM or TCM-199, in the in-vitro culture of equine preantral follicles. The research found promising initial results suggesting that the use of α-MEM for 7 days can promote preantral follicle development in cultures obtained by the Biopsy Pick-Up method.
Methodology
- The research used equine ovarian tissue obtained from ten mares via the Biopsy Pick-Up method.
- These tissues were immediately subjected to a histological analysis, or they were cultured for a short period of either one day (D1) or seven days (D7).
- Two types of media were used for the culture: α-MEM and TCM-199, creating five treatment groups for the experiment. These groups were; noncultured control, α-MEM:D1, TCM-199:D1, α-MEM:D7, and TCM-199:D7.
Follicle Analysis
- The researchers evaluated the preantral follicles based on their class (primordial, transitional, primary, secondary) and their morphology (normal vs. abnormal).
- In total, 142 preantral follicles were analyzed for this study.
Findings
- No follicles were observed in the TCM-199:D7 treatment group, suggesting this method was unsuccessful.
- There was a higher proportion of primordial follicles in the control group compared to the α-MEM culture of 7 days. However, there were more of the primary (developing) follicles in the α-MEM:D7 group than in the control, indicative of development in vitro.
- The study also found a higher percentage of morphologically normal follicles in the α-MEM:D1 treatment group compared to the TCM-199:D1 treatment group, implying that α-MEM might be a better medium for follicle development in vitro.
- In terms of growth and development, the mean follicle and oocyte diameters were larger in the α-MEM:D1 and α-MEM:D7 groups compared to the control group. This suggests that the α-MEM medium can promote growth.
Conclusion
- Based on these results, the research concludes that utilising α-MEM for in-vitro culture of equine ovarian fragments helps to promote preantral follicle development. This discovery could have implications for advancements in reproductive biotechnology in horses.
Cite This Article
APA
Haag KT, Magalhães-Padilha DM, Fonseca GR, Wischral A, Gastal MO, King SS, Jones KL, Figueiredo JR, Gastal EL.
(2013).
In vitro culture of equine preantral follicles obtained via the Biopsy Pick-Up method.
Theriogenology, 79(6), 911-917.
https://doi.org/10.1016/j.theriogenology.2013.01.001 Publication
Researcher Affiliations
- Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, Illinois, USA.
MeSH Terms
- Animals
- Biopsy / methods
- Biopsy / veterinary
- Cell Culture Techniques / veterinary
- Culture Media
- Female
- Horses / physiology
- Ovarian Follicle / cytology
- Ovarian Follicle / growth & development
Citations
This article has been cited 7 times.- Taghizabet N, Bahmanpour S, Fard NZ, Rezaei-Tazangi F, Hassanpour A, Nejad EK, Aliakbari F, Dehghani F. In vitro growth of the ovarian follicle: taking stock of advances in research. JBRA Assist Reprod 2022 Aug 4;26(3):508-521.
- Souza SS, Alves BG, Alves KA, Brandão FAS, Brito DCC, Gastal MO, Rodrigues APR, Figueireod JR, Teixeira DIA, Gastal EL. Heterotopic autotransplantation of ovarian tissue in a large animal model: Effects of cooling and VEGF. PLoS One 2020;15(11):e0241442.
- 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.
- Castañeda OJR, de Aguiar FLN, de Sá NAR, Morais MLGDS, Cibin FWS, Torres CAA, de Figueiredo JR. Powdered coconut water (ACP 406®) as an alternative base culture medium for in vitro culture of goat preantral follicles enclosed in ovarian tissue. Anim Reprod 2019 Nov 18;16(4):838-845.
- Max MC, Bizarro-Silva C, Búfalo I, González SM, Lindquist AG, Gomes RG, Barreiros TRR, Lisboa LA, Morotti F, Seneda MM. In vitro culture supplementation of EGF for improving the survival of equine preantral follicles. In Vitro Cell Dev Biol Anim 2018 Dec;54(10):687-691.
- Alves KA, Alves BG, Gastal GD, de Tarso SG, Gastal MO, Figueiredo JR, Gambarini ML, Gastal EL. The Mare Model to Study the Effects of Ovarian Dynamics on Preantral Follicle Features. PLoS One 2016;11(2):e0149693.
- 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.
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