Effect of LH and Estradiol-17β Supplementation at Different Time Points on In vitro Development of Preantral Follicles in Sheep

2020 ◽  
Author(s):  
L.S.S. Varaprasad Reddy ◽  
B.R. Naik ◽  
A.V.N. Sivakumar ◽  
B. Punyakumari ◽  
J. Suresh
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Follicular Development ◽  
Standard Medium ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Time Points ◽  
Vitro Development ◽  
Estradiol 17Β

Background: Ovarian follicular development and growth are controlled by many hormones and growth factors. Despite the fact that LH and estradiol-17β have been utilized for the in vitro culture of preantral follicles yet, the suitable time points of supplementation of LH and estradiol-17β is not known. Therefore this study aimed to investigate the influence of addition of LH and estradiol-17β at different time points on in vitro development of preantral follicles (PFs’) in sheep. Method: Preantral follicles isolated from the ovarian cortical slices using micro dissection method were cultured for six days in Bicarbonate buffered Tissue culture medium 199B (TCM 199B) or in a standard culture medium supplemented with LH (2 μg/ml) and estradiol-17β (5 ng/ml) at different points during the culture period. COCs isolated from the follicles at the end of six day culture in different treatments were subjected to in vitro maturation for additional 24h. Result: Supplementation of LH and estradiol-17β during last two days of the culture supported better proportion of PFs’ exhibiting growth whereas supplementation of LH and estradiol-17β during first two days of the culture supported better average increase in diameter and proportion of PFs’ exhibiting antrum formation at the end of six day culture. Further the oocytes in COCs isolated at the end of culture in these treatments and subsequently subjected to in vitro maturation (IVM) for 24hr developed at a higher frequency to MII (metaphase II) stage. Supplementation of LH and estradiol-17β to TCM 199B culture medium in early stages followed by standard medium alone in later stages supports better development of PFs’ in vitro. Following supplementation with LH and estradiol-17β for the first two days culture of PFs’ in standard medium appears to be advantageous for the development of preantral follicles in vitro.

Bovine dominant follicular fluid promotes the in vitro development of goat preantral follicles

2012 ◽  
Vol 24 (3) ◽  
pp. 490 ◽  
Author(s):  
A. B. G. Duarte ◽  
V. R. Araújo ◽  
R. N. Chaves ◽  
G. M. Silva ◽  
D. M. Magalhães-Padilha ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Culture Medium ◽  
Follicular Development ◽  
Ultrastructural Analysis ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Nuclear Membranes ◽  
Vitro Development ◽  
First Time

The aim of this study was to evaluate the effect of follicular fluid collected from bovine dominant follicles (bFF) on the in vitro development of goat preantral follicles and determine the best time to add this supplement to the culture medium. The preantral follicles were isolated and randomly distributed into four treatments in absence (control) or presence of 10% of bFF added on Days 0 (FF0–18), 6 (FF6–18) or 12 (FF12–18) of culture onwards. After 18 days, follicular development was assessed based on follicular survival, antral cavity formation, increased follicular diameter as well as fully grown oocyte (>110 μm) viability and meiosis resumption. The oocytes from the cultured follicles were in vitro-matured and processed for fluorescence or ultrastructural analysis. The results showed that on Day 18 the treatment FF0–18 had a significantly higher (P < 0.05) survival than control and FF12–18, but not FF6–18. The addition of bFF at the beginning of culture (FF0–18 and FF6–18) promoted a high percentage of follicular growth, meiosis resumption and early antrum formation. Moreover, this study described for the first time the ultrastructural analysis of caprine oocytes grown in vitro. This evaluation revealed that in the presence of bFF on (FF0–18) the in vitro-grown oocytes presented normal organelle distribution and well-defined, intact plasma and nuclear membranes. In conclusion, bFF originating from dominant follicles maintain the survival and promote the in vitro growth of goat preantral follicles when added at the beginning of culture.

scholarly journals The effects of supplemented sericin on in vitro maturation and preimplantation development of mouse embryos: An experimental study

2021 ◽  
pp. 921-928
Author(s):  
Omid Banafshi ◽  
Sherko Nasseri ◽  
Leila Farhadi ◽  
Masoud Alasvand ◽  
Mohammad Bagher Khadem-Erfan ◽  
...  
Keyword(s):  
Embryo Culture ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Control Group ◽  
Vitro Fertilization ◽  
In Vitro Development ◽  
Embryo Culture Medium ◽  
Vitro Development

Background: Mouse embryo culture condition is an essential part of transgenic, reproductive and developmental biology laboratories. Mouse embryonic culture media may have a high risk of serum contamination with pathogens.  Objective: To investigate the effect of sericin as an embryo culture medium supplement on in vitro maturation (IVM), in vitro fertilization (IVF), and development of the preimplantation embryo in mice. Materials and Methods: The effects of sericin at three concentrations (subgroups) of 0.1%, 0.5%, and 1% as a medium supplement on IVM, IVF, and in vitro development of mouse embryos were separately investigated and compared with a sericin-free (control) group. The cumulative effect of the three concentrations was evaluated for IVM + in vitro development and IVF + in vitro development as follow-up groups. Results: In the IVM group, compared to the control group, the number of oocysts reaching the MII stage was significantly higher when 1% sericin was used (161/208 = 77.4%). No significant results were observed in the IVF and in vitro development groups with different concentrations of sericin compared to the control group. Among the follow-up groups, in the IVM + in vitro development group, the number of oocytes was higher after passing the IVM and IVF and reaching the blastocysts stage when 1% sericin was used, compared with other sericin subgroups. A significant difference was also noted when compared with the control group (p = 0.048). The IVF + in vitro development study group, on the other hand, did not show any significant relationship. Conclusion: It can be concluded that 1% sericin can be used as a supplement in mouse embryo cultures to improve the IVM rate. Also, based on the findings, sericin appears to be an effective supplement which can have a positive effect on the development of embryos derived from IVM. Key words: Sericin, In vitro maturation, In vitro fertilization, Preimplantation embryo, Culture medium, Mice.

Effect of alpha lipoic acid on in vitro development of bovine secondary preantral follicles

2017 ◽  
Vol 88 ◽  
pp. 124-130 ◽  
Author(s):  
Khairy M.A. Zoheir ◽  
Gamaleldin I. Harisa ◽  
Ahmed A. Allam ◽  
Liguo Yang ◽  
Xiang Li ◽  
...  
Keyword(s):  
Lipoic Acid ◽  
Alpha Lipoic Acid ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Vitro Development

scholarly journals Effect of Melatonin on the In Vitro Maturation of Porcine Oocytes, Development of Parthenogenetically Activated Embryos, and Expression of Genes Related to the Oocyte Developmental Capability

Animals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 209 ◽  
Author(s):  
Ling Yang ◽  
Qingkai Wang ◽  
Maosheng Cui ◽  
Qianjun Li ◽  
Shuqin Mu ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Polar Body ◽  
Melatonin Receptors ◽  
Melatonin Treatment ◽  
In Vitro Development ◽  
First Polar Body ◽  
Mitochondrial Distribution ◽  
Polar Body Extrusion ◽  
Vitro Development

Melatonin treatment can improve quality and in vitro development of porcine oocytes, but the mechanism of improving quality and developmental competence is not fully understood. In this study, porcine cumulus–oocyte complexes were cultured in TCM199 medium with non-treated (control), 10−5 M luzindole (melatonin receptor antagonist), 10−5 M melatonin, and melatonin + luzindole during in vitro maturation, and parthenogenetically activated (PA) embryos were treated with nothing (control), or 10−5 M melatonin. Cumulus oophorus expansion, oocyte survival rate, first polar body extrusion rate, mitochondrial distribution, and intracellular levels of reactive oxygen species (ROS) and glutathione of oocytes, and cleavage rate and blastocyst rate of the PA embryos were assessed. In addition, expression of growth differentiation factor 9 (GDF9), tumor protein p53 (P53), BCL2 associated X protein (BAX), catalase (CAT), and bone morphogenetic protein 15 (BMP15) were analyzed by real-time quantitative PCR. The results revealed that melatonin treatment not only improved the first polar body extrusion rate and cumulus expansion of oocytes via melatonin receptors, but also enhanced the rates of cleavage and blastocyst formation of PA embryos. Additionally, melatonin treatment significantly increased intraooplasmic level of glutathione independently of melatonin receptors. Furthermore, melatonin supplementation not only significantly enhanced mitochondrial distribution and relative abundances of BMP15 and CAT mRNA, but also decreased intracellular level of ROS and relative abundances of P53 and BAX mRNA of the oocytes. In conclusion, melatonin enhanced the quality and in vitro development of porcine oocytes, which may be related to antioxidant and anti-apoptotic mechanisms.

scholarly journals Kit ligand and insulin-like growth factor I affect the in vitro development of ovine preantral follicles

2013 ◽  
Vol 115 (1-3) ◽  
pp. 99-102 ◽  
Author(s):  
V.B. Luz ◽  
V.R. Araújo ◽  
A.B.G. Duarte ◽  
G.M. Silva ◽  
R.N. Chaves ◽  
...  
Keyword(s):  
Growth Factor ◽  
Insulin Like Growth Factor ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Factor I ◽  
Kit Ligand ◽  
Growth Factor I ◽  
Vitro Development
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