scholarly journals Effect of MEM Vitamins Supplementation of In vitro Maturation Medium and In vitro Culture Medium on the Development of Porcine Embryos

2011 ◽  
Vol 24 (11) ◽  
pp. 1541-1546
Author(s):  
J. Y. Kim ◽  
E. J. Lee ◽  
J. M. Park ◽  
H. D. Park
Keyword(s):  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Maturation Medium

Cysteamine supplementation of in vitro maturation medium, in vitro culture medium or both media promotes in vitro development of buffalo (Bubalus bubalis) embryos

2008 ◽  
Vol 20 (2) ◽  
pp. 253 ◽  
Author(s):  
T. Anand ◽  
D. Kumar ◽  
M. S. Chauhan ◽  
R. S. Manik ◽  
P. Palta
Keyword(s):  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Bubalus Bubalis ◽  
Cell Number ◽  
Hatching Rate ◽  
Cleavage Rate ◽  
Maturation Medium ◽  
Vitro Development

The effects of supplementation of in vitro maturation (IVM) or in vitro culture (IVC) or both IVM and IVC media with cysteamine on the yield, hatching rate (HR) and total cell number (TCN) of buffalo blastocysts were examined. Oocytes obtained from slaughterhouse buffalo ovaries were subjected to IVM and IVF. The IVM or IVC media were supplemented with 0, 50, 100 or 200 µm cysteamine. Supplementation of IVM medium with 50 µm cysteamine increased (P < 0.01) the cleavage rate and blastocyst yield without affecting the HR and TCN whereas a higher concentration of 200 µm significantly (P < 0.05) reduced the blastocyst yield but not TCN. Similar increases in blastocyst yield, without any effect on HR and TCN were observed after supplementation of the IVC medium with 100 (P < 0.01) or 50 µm (P < 0.05) cysteamine, whereas 200 µm cysteamine was ineffective. Supplementation of both IVM medium with 50 µm cysteamine and of IVC medium with 100 µm cysteamine increased the yield of blastocysts and hatched blastocyst by over 100% (P < 0.01) compared with the controls without any adverse effects on HR or TCN. The results of the present study suggest that supplementation of both IVM and IVC media improves the yield of blastocysts without compromising their health.

148 Effect of concentration of methionine in maturation and culture medium on cleavage rate of oocytes from alpaca (Lama pacos)

2019 ◽  
Vol 31 (1) ◽  
pp. 199
Author(s):  
M. L. Uchuari ◽  
M. Artica ◽  
J. C. Villanueva ◽  
W. F. Huanca ◽  
W. Huanca
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Egg Yolk ◽  
Cumulus Cells ◽  
Cleavage Rate ◽  
Sodium Pyruvate ◽  
Maturation Medium ◽  
Thermos Flask ◽  
Lama Pacos

Maturation time of oocytes from alpacas is around 38 to 40h (Huanca et al. 2009) that would induce an increase in reactive oxygen species during in vitro maturation and IVF and cause cytotoxic damage to gametes. The objective of this study was to determine the optimal concentration of methionine during in vitro maturation on cleavage rate of alpacas oocytes following IVF. Cumulus-oocyte complexes were collected from slaughterhouse ovaries and transported in a thermos flask containing a saline solution 0.9% and antibiotic, antimycotic at 35°C. Cumulus-oocyte complexes were aspirated from follicles &gt;2mm and evaluated with a stereomicroscope for selection. Only cumulus-oocyte complexes with a homogeneous cytoplasm and with 2 or more layers of cumulus cells were selected to be cultured in maturation medium TCM-199 supplemented with 10% FCS (v:v) plus 0.5μg mL−1 FSH, 10μg mL−1 hCG, 0.2mM sodium pyruvate, 50μg mL−1gentamycin and 1μg mL−1 oestradiol under mineral oil by 38h. Testes of mature males were collected from a slaughterhouse and transported to the laboratory. Caudal epididymide was isolated, and fluid, rich in spermatozoa, was aspirated in syringes containing 2mL of Tris-fructose-egg yolk extender. Motile spermatozoa were obtained by centrifugation at 700×g in a Percoll discontinuous gradient (22.5: 45.0%) for 10min. The supernatant was removed by aspiration, and the pellet was resuspended in TL stock and centrifuged again at 700×g for 5min. Spermatozoa and oocytes were co-incubated by 18h at 39°C with 5% CO2. Presumptive zygotes were culture in KSOMaa medium and evaluated at 72h. The treatments include 0, 14 and 21 μM of methionine in maturation and culture medium. Data were analysed by ANOVA, and results are presented in Table 1. The results suggest that addition of methionine in maturation and culture medium improve the cleavage rate in oocytes from alpacas. Table 1.Cleavage rate (%) following in vitro maturation at different concentrations of methionine Proyect 405-PNICP-PIAP-2014, INNOVATE-PERU, is acknowledged.

scholarly journals Applying the nanobiotechnological methods for optimize the in vitro cultivation technology for cows oocytes

2018 ◽  
Vol 22 ◽  
pp. 257-261 ◽  
Author(s):  
S. I. Kovtun ◽  
A. B. Zyuzyun ◽  
O. V. Shcherbak ◽  
P. A. Trotskiy
Keyword(s):  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
In Vitro Cultivation ◽  
Vitro Fertilization ◽  
Cultivation Technology ◽  
And Control ◽  
Positive Effect ◽  
Generative Cells

Aim. Investigated effect nanomaterial of highly dispersed ultra fine silica (UFS) by carbohydrate – sucrose (UFS/sucrose) on the effectiveness of meiotic maturation cows oocytes in vitro. Methods. The fresh and frozen – thawed cow oocyte-cumulus complex (OCC) was divided into four groups: three experimental in which the cultivation was carried out in a medium containing of 0, 1; 0, 01 and 0,001 % UFS/sucrose and control without adding nanomaterial. Results. It was concluded that the addition of UFS/sucrose in 0.001 % concentration is effective for elevation level of oocytes maturation and provides reception of 76,8% oocytes that induced the metaphase II of meiosis. Adding carbohydrate – sucrose (UFS/sucrose) in 0.001 % concentration to the culture medium frozen – thawed cow generative cells, make positive effect on in vitro fertilization and provide embryos quantity enhancement to 33.3 %. Conclusions. Addition of UFS/sucrose in 0.001 % concentration to the culture medium have increase effect and promote level in vitro maturation of cows oocytes rising to 76.8 %. Usage of UFS/sucrose in 0.001 % concentration as part of in vitro culture medium for cows oocyte-cumulus complex conduce rising quantity of cattle embryos to 33.3 % after in vitro fecundation frozen – thawed and maturation oocytes. Keywords: oocytes, in vitro culture, embryos, nanomaterial, ultra fine silica (UFS).

Anethole improves blastocysts rates together with antioxidant capacity when added during bovine embryo culture rather than in the in vitro maturation medium

Zygote ◽  
2019 ◽  
Vol 27 (6) ◽  
pp. 382-385 ◽  
Author(s):  
J.C. Anjos ◽  
F.L.N. Aguiar ◽  
N.A.R. Sá ◽  
J.F. Souza ◽  
F.W.S. Cibin ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Cumulus Cells ◽  
Bovine Embryo ◽  
Oxygen Species ◽  
Embryo Production ◽  
Maturation Medium ◽  
Bovine Oocytes ◽  
Medium Supplementation

SummaryWe performed the exposure of bovine oocytes to anethole during in vitro maturation (0 or 300 µg/ml), during in vitro embryo production (0, 30, 300 or 2000 µg/ml), or during both periods to determine the rates of 2−4 cells embryos, blastocysts rates and cells numbers, as well as the production of reactive oxygen species (ROS). Bovine ovaries (n = 240) were collected from a local abattoir after slaughter and cumulus–oocyte complexes (COCs) with homogeneous and non-dark cytoplasm, surrounded by two or more compact layers of cumulus cells, and an intact zona pellucida were selected for in vitro maturatuion (IVM). Mature oocytes were then submitted to in vitro fertilization (IVF) and in vitro embryo production (IVP) in culture medium supplemented or not with different concentrations of anethole, as described above. Although IVM medium supplementation with 300 µg/ml anethole improved the rates of bovine blastocysts formation, we demonstrated that IVP medium supplementation with 30 µg/ml anethole, regardless of IVM medium enrichment, considerably enhanced blastocysts rates. Furthermore, ROS levels were decreased only when anethole was added to the IVP medium without previous IVM medium supplementation.

scholarly journals Supplementation of Follicle Stimulating Hormon Into In vitro Maturation Medium to Increase Oocytes Maturation and 4 Cell Stadium Embryo Development of Bligon Goat

2018 ◽  
Vol 42 (2) ◽  
Author(s):  
Yanuar Achadri ◽  
Sigit Bintara ◽  
Diah Tri Widayati
Keyword(s):  
Tissue Culture ◽  
Embryo Development ◽  
Embryo Culture ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Tissue Culture Medium ◽  
Cleavage Rate ◽  
Maturation Medium ◽  
Phosphate Buffered Saline

The study was carried out to investigate the effect of follicle stimulating hormon (FSH) into in vitro maturation medium to increase oocytes maturation and 4 cell stadium embryo development of Bligon goat. Goat ovaries were obtained from a slaughterhouse and transported to the laboratory in a flask of NaCl at temperature of 31 – 34°C. Oocytes were aspirated from 2 – 6 mm of follicles into a 3 mL syringe (23G needle) that contained Dulbecco’s Phosphate-Buffered Saline. Oocytes were divided into three groups, i.e tissue culture medium (TCM) with FSH supplementation 0, 50, and 100 IU/mL. Oocytes were put into those medium and incubated on 39°C, 5% CO2, and 95% humidity for 24 hours. Matured oocytes were fertilized with capacitated frozen thawed-semen and incubated on 39°C, 5% CO2, and 95% humidity for 5 hours. Fertilized oocytes were washed for 3 times in TCM and incubated in the same condition for embryo culture. The data of FSH supplementation and embryo development were analyzed using randomized completely one way classification. The results showed that the percentages of mature oocytes from FSH supplementation 0, 50, and 100 IU/mL were 70,48±23,22, 78,48±15,80, and 80,29±12,86%, respectively. Cleavage rate of the two cells stage were 36,00±14,22, 44,00±33,94, and 57,45±31,78%, respectively, and for the 4 cells stage were 27,33±22,04, 35,33±40,73, and 39,45±20,38%. It is concluded that supplementation of FSH in the maturation medium could not increase the percentages of in vitro maturation and embryo development.

scholarly journals Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Average Pore Size ◽  
Physiological Responses ◽  
Gum Arabic ◽  
Shoot Elongation ◽  
Hydroxyethyl Cellulose ◽  
Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

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