Blastocyst development in equine oocytes with low meiotic competence after suppression of meiosis with roscovitine prior to in vitro maturation.
Abstract: This study was conducted to evaluate the in vitro development of equine oocytes with compact cumuli that had been subjected to a period of meiotic suppression with roscovitine before in vitro maturation. In experiment 1, oocytes were recovered from slaughterhouse-derived ovaries and held in M199 + 10% fetal bovine serum containing 66 microM roscovitine with or without an overlay of mineral oil in 5% CO2 in air at 38.2 degrees C for 16-18 or 24 h. No oocytes treated with roscovitine in the absence of an oil overlay for 16-18 h were maturing, compared with 2-4% of oocytes in other treatments. In experiment 2, oocytes were either fixed immediately after recovery, or were cultured for 18 h in the presence or absence of roscovitine. Oocytes cultured in the absence of roscovitine had a significantly higher rate of meiotic resumption (18%) than was found in the other two treatments (0). In experiment 3, oocytes were matured immediately or after 16-18 h culture with roscovitine. Maturation rates were similar between oocytes previously treated with roscovitine (22%) and control oocytes (19%). Mature oocytes were fertilized by intracytoplasmic sperm injection and then cultured, with or without oviductal epithelial cells, for 7.5 days. There was no significant effect of roscovitine treatment on blastocyst development. Development to blastocyst of roscovitine-treated oocytes in DMEM/F-12 + co-culture (37%) was significantly higher than that of control oocytes in DMEM/F-12 without co-culture (14%). These data indicate that equine oocytes with compact cumuli can be held in roscovitine before maturation without any harmful effect on blastocyst formation.
Publication Date: 2006-05-17 PubMed ID: 16700969DOI: 10.1017/S0967199406003534Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study investigates the in-vitro development of horse oocytes that had temporary meiotic suppression using roscovitine before the process of in-vitro maturation. The findings suggest, meiotic suppression with roscovitine didn’t have any harmful impact on the formation of blastocysts.
Research Methods and Process
- The research carried out three different experiments to understand the effect of roscovitine on oocytes, which are immature egg cells. The study used equine oocytes possessing compact cumuli, structures vital for the development of the egg.
- In the first experiment, the oocytes were recovered from ovaries and then exposed to a culture medium with roscovitine. The presence or absence of an overlay of mineral oil and different incubation periods were also assessed.
- In the second experiment, oocytes were observed right after recovery or were cultured with or without roscovitine for 18 hours and compared.
- In the third experiment, oocytes were immediately matured after 16-18 hours of culture with roscovitine or without roscovitine. These matured oocytes were fertilized by the process of intracytoplasmic sperm injection and cultured for 7.5 days.
Key Findings
- The observations from the first experiment showed that no oocytes matured when treated with roscovitine without an oil overlay for 16-18 hours, compared to 2-4% of oocytes in other methods.
- The second experiment revealed that the oocytes cultured without roscovitine had a significantly higher rate of meiotic resumption, which was 18%, than the other two treatments, which recorded 0%.
- The third experiment noted that the maturation rates were insignificantly different between the oocytes previously treated with roscovitine and the control oocytes. Also, regardless of the roscovitine treatment, it had no significant effect on blastocyst (an early stage in the development of an embryo) development.
- Interestingly, the roscovitine-treated oocytes that were co-cultured in DMEM/F-12 (a type of cell culture medium) showed a significantly higher blastocyst development resting at 37% compared to control oocytes without co-culturing that led to only 14% development.
- The research carried out three different experiments to understand the effect of roscovitine on oocytes, which are immature egg cells. The study used equine oocytes possessing compact cumuli, structures vital for the development of the egg.
- In the first experiment, the oocytes were recovered from ovaries and then exposed to a culture medium with roscovitine. The presence or absence of an overlay of mineral oil and different incubation periods were also assessed.
- In the second experiment, oocytes were observed right after recovery or were cultured with or without roscovitine for 18 hours and compared.
- In the third experiment, oocytes were immediately matured after 16-18 hours of culture with roscovitine or without roscovitine. These matured oocytes were fertilized by the process of intracytoplasmic sperm injection and cultured for 7.5 days.
Key Findings
- The observations from the first experiment showed that no oocytes matured when treated with roscovitine without an oil overlay for 16-18 hours, compared to 2-4% of oocytes in other methods.
- The second experiment revealed that the oocytes cultured without roscovitine had a significantly higher rate of meiotic resumption, which was 18%, than the other two treatments, which recorded 0%.
- The third experiment noted that the maturation rates were insignificantly different between the oocytes previously treated with roscovitine and the control oocytes. Also, regardless of the roscovitine treatment, it had no significant effect on blastocyst (an early stage in the development of an embryo) development.
- Interestingly, the roscovitine-treated oocytes that were co-cultured in DMEM/F-12 (a type of cell culture medium) showed a significantly higher blastocyst development resting at 37% compared to control oocytes without co-culturing that led to only 14% development.
- The observations from the first experiment showed that no oocytes matured when treated with roscovitine without an oil overlay for 16-18 hours, compared to 2-4% of oocytes in other methods.
- The second experiment revealed that the oocytes cultured without roscovitine had a significantly higher rate of meiotic resumption, which was 18%, than the other two treatments, which recorded 0%.
- The third experiment noted that the maturation rates were insignificantly different between the oocytes previously treated with roscovitine and the control oocytes. Also, regardless of the roscovitine treatment, it had no significant effect on blastocyst (an early stage in the development of an embryo) development.
- Interestingly, the roscovitine-treated oocytes that were co-cultured in DMEM/F-12 (a type of cell culture medium) showed a significantly higher blastocyst development resting at 37% compared to control oocytes without co-culturing that led to only 14% development.
The overall research suggests that the equine oocytes with compact cumuli can undergo a holding period in roscovitine before maturation, indicating it has no harmful influence on blastocyst formation. The better development in roscovitine-treated oocytes when co-cultured, indicates a potentially beneficial interaction not yet fully understood.
Cite This Article
APA
Choi YH, Love LB, Varner DD, Hinrichs K.
(2006).
Blastocyst development in equine oocytes with low meiotic competence after suppression of meiosis with roscovitine prior to in vitro maturation.
Zygote, 14(1), 1-8.
https://doi.org/10.1017/S0967199406003534 Publication
Researcher Affiliations
- Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843-4466, USA.
MeSH Terms
- Animals
- Blastocyst / drug effects
- Blastocyst / physiology
- Coculture Techniques
- Epithelial Cells / cytology
- Epithelial Cells / physiology
- Fallopian Tubes / cytology
- Fallopian Tubes / physiology
- Female
- Horses / embryology
- Meiosis / drug effects
- Oocytes / cytology
- Oocytes / drug effects
- Oocytes / growth & development
- Protein Kinase Inhibitors / metabolism
- Protein Kinase Inhibitors / pharmacology
- Purines / metabolism
- Purines / pharmacology
- Roscovitine
- Sperm Injections, Intracytoplasmic
Citations
This article has been cited 4 times.- Cortez JV, Hardwicke K, Méndez-Calderón CE, Grupen CG. Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals. Animals (Basel) 2025 Jul 3;15(13).
- Fakhar-I-Adil M, Angel-Velez D, Araftpoor E, Amin QA, Hedia M, Bühler M, Gevaert K, Menten B, Van Soom A, Chuva de Sousa Lopes SM, Stoop D, De Roo C, Smits K, Heindryckx B. Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development Without Inducing Genetic Aberrations. Int J Mol Sci 2025 Jun 8;26(12).
- Luo Y, Li J, Zheng L, Reyimjan Y, Ma Y, Huang S, Liu H, Zhou G, Bai J, Zhu Y, Sun Y, Zou X, Hou Y, Fu X. Procyanidin B2 improves developmental capacity of bovine oocytes via promoting PPARγ/UCP1-mediated uncoupling lipid catabolism during in vitro maturation. Cell Prolif 2024 Nov;57(11):e13687.
- Farghaly T, Khalifa E, Mostafa S, Hussein M, Bedaiwy M, Ahmady A. The effect of temporary meiotic attenuation on the in vitro maturation outcome of bovine oocytes. In Vitro Cell Dev Biol Anim 2015 Aug;51(7):662-71.
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