Effects of follicular fluid supplementation of in-vitro maturation medium on the fertilization and development of equine oocytes after in-vitro fertilization or intracytoplasmic sperm injection.
Abstract: The aim of this study was to compare the effect of the addition of follicular fluid (FF) collected from preovulatory follicles with that of oestrous mare serum (EMS) (acting as the control) to TCM-199 medium on the in-vitro maturation, fertilization and development of equine cumulus-enclosed oocytes. Oocytes (<30 mm in diameter) were obtained from the ovaries of slaughtered mares. After in-vitro maturation in the presence of the two supplements, their fertilization, cleavage and developmental potential were compared after conventional in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) using frozen-thawed spermatozoa. Follicular fluid did not increase the maturation of oocytes to metaphase II stage compared to control. After IVF, there was no difference in fertilization rates between FF-supplemented oocytes and controls (7/87, 8.4% of oocytes showing two pronuclei with FF versus 7/116, 6% with EMS; not significant). However, after ICSI, FF-supplemented oocytes showed significantly increased normal fertilization (32/85, 37.6% of two-pronuclear oocytes) and developmental potential (15/31, 48% cleavage) compared to the control oocytes (7/47, 14.9%, P < 0.01; and 2/48, 4%, P < 0.01, respectively). Overall, ICSI resulted in increased fertilization rates compared to IVF, regardless of the presence or absence of FF (39/132, 29.5% with ICSI versus 14/203, 6.9%). These results suggest that follicular fluid supplementation may improve the maturity of equine cumulus-enclosed oocytes sufficiently for the successful use of ICSI, but not sufficiently for normal sperm-egg interaction occurring during IVF.
Publication Date: 1998-02-10 PubMed ID: 9455850DOI: 10.1093/humrep/12.12.2766Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the impact of adding follicular fluid (FF) from preovulatory follicles to the medium used for in-vitro maturation of horse oocytes on their development and fertilization. The study found that while adding FF did not lead to greater maturity of oocytes for regular in-vitro fertilization (IVF) methods, it significantly improved oocyte maturity for intracytoplasmic sperm injection (ICSI), a variant of traditional IVF.
Study Purpose and Method
- The study aimed to ascertain the effect of adding follicular fluid, taken from preovulatory follicles, on the in-vitro maturation process of horse oocytes. The end goal was to see if this modification impacts their ability to fertilize and develop.
- Oocytes, or immature egg cells, were collected from the ovaries of slaughtered mares and matured in-vitro using FF and oestrous mare serum (EMS) to complement the TCM-199 medium used for this process.
- Frozen-thawed spermatozoa were then used to fertilize the oocytes either through the conventional in-vitro fertilization technique or intracytoplasmic sperm injection.
Research Findings
- The addition of FF did not enhance the maturity of oocytes to a greater degree compared to the control oocytes which only had EMS for the in-vitro maturation process.
- There was no discernible difference in fertilization rates between oocytes that had been matured through FF supplementation and those that didn’t. This conclusion held true for the traditional in-vitro fertilization technique.
- However, when fertilized through ICSI, oocytes that had been matured in FF exhibited significantly increased normal fertilization and developmental potential than the control group.
- The study also noted that regardless of whether FF was present or not, ICSI generally resulted in higher fertilization rates than the traditional IVF process.
Implications of the Study
- The results suggest that adding follicular fluid during the in-vitro maturation process could boost the maturity of horse oocytes enough to have successful ICSI, but not enough for normal sperm-egg interaction occurring during traditional IVF.
Cite This Article
APA
Dell'Aquila ME, Cho YS, Minoia P, Traina V, Lacalandra GM, Maritato F.
(1998).
Effects of follicular fluid supplementation of in-vitro maturation medium on the fertilization and development of equine oocytes after in-vitro fertilization or intracytoplasmic sperm injection.
Hum Reprod, 12(12), 2766-2772.
https://doi.org/10.1093/humrep/12.12.2766 Publication
Researcher Affiliations
- Institute of Reproductive Biology and Obstetrics, Faculty of Veterinary Medicine, University of Bari, Italy.
MeSH Terms
- Animals
- Culture Media
- Culture Techniques
- Embryonic and Fetal Development
- Female
- Fertilization in Vitro
- Follicular Fluid / physiology
- Horses
- Microinjections
- Oocytes / physiology
Citations
This article has been cited 13 times.- Samaniego JX, Pesantez JL, Ayala LE, Perea FP, Galarza DA, Dutan JB, Ruiz S. Effects of Follicular Fluid and Serum Supplementation on Cumulus Cell Expansion and Nuclear Progression of Guinea Pig Oocytes, Using a Baseline Medium Established with Bovine Oocytes. Animals (Basel) 2025 Feb 25;15(5).
- Negrón-Pérez VM, Al Naib A, Zezeski AL, McCracken-Harlow VL, Perry GA, Ealy AD, Rhoads ML. Cumulus cell expansion, nuclear maturation and embryonic development of bovine cumulus-oocyte complexes matured in varying concentrations of follicular fluid. PLoS One 2025;20(2):e0318376.
- Aoki S, Inoue Y, Hara S, Itou J, Shirasuna K, Iwata H. microRNAs associated with the quality of follicular fluids affect oocyte and early embryonic development. Reprod Med Biol 2024 Jan-Dec;23(1):e12559.
- Dipali SS, Suebthawinkul C, Burdette JE, Pavone ME, Duncan FE. Human follicular fluid elicits select dose- and age-dependent effects on mouse oocytes and cumulus-oocyte complexes in a heterologous in vitro maturation assay. Mol Hum Reprod 2023 Nov 1;29(11).
- Maitan PP, Bromfield EG, Hoogendijk R, Leung MR, Zeev-Ben-Mordehai T, van de Lest CH, Jansen JWA, Leemans B, Guimarães JD, Stout TAE, Gadella BM, Henning H. Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa. Front Cell Dev Biol 2021;9:772254.
- Tharasanit T, Thuwanut P. Oocyte Cryopreservation in Domestic Animals and Humans: Principles, Techniques and Updated Outcomes. Animals (Basel) 2021 Oct 13;11(10).
- Satué K, Fazio E, Medica P. Can the Presence of Ovarian Corpus Luteum Modify the Hormonal Composition of Follicular Fluid in Mares?. Animals (Basel) 2020 Apr 9;10(4).
- Leemans B, Stout TAE, Soom AV, Gadella BM. pH-dependent effects of procaine on equine gamete activation†. Biol Reprod 2019 Nov 21;101(5):1056-1074.
- Ahamad SR, Yaqoob SH, Khan A, Shakeel F. Metabolite profile and elemental determination of camel follicular fluid by GC-MS and ICP-MS. Trop Anim Health Prod 2019 Nov;51(8):2447-2454.
- Yuan Y, Spate LD, Redel BK, Tian Y, Zhou J, Prather RS, Roberts RM. Quadrupling efficiency in production of genetically modified pigs through improved oocyte maturation. Proc Natl Acad Sci U S A 2017 Jul 18;114(29):E5796-E5804.
- Abdoon AS, Abdel-Rahman HA, Shawki SM, Kandil OM, Fathalla SI. Influence of follicle size, methods of retrieval on oocytes yield and morphology in Egyptian Jennies ovaries with special reference to maturation rate in vitro. Vet Res Commun 2014 Dec;38(4):287-95.
- McPartlin LA, Visconti PE, Bedford-Guaus SJ. Guanine-nucleotide exchange factors (RAPGEF3/RAPGEF4) induce sperm membrane depolarization and acrosomal exocytosis in capacitated stallion sperm. Biol Reprod 2011 Jul;85(1):179-88.
- Lange Consiglio A, Dell'Aquila ME, Fiandanese N, Ambruosi B, Cho YS, Bosi G, Arrighi S, Lacalandra GM, Cremonesi F. Effects of leptin on in vitro maturation, fertilization and embryonic cleavage after ICSI and early developmental expression of leptin (Ob) and leptin receptor (ObR) proteins in the horse. Reprod Biol Endocrinol 2009 Oct 16;7:113.
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