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).

286 INFLUENCE OF REVERSIBLE MEIOSIS INHIBITION ON SWINE EMBRYOS PRODUCED BY IN VITRO FERTILIZATION AND PATHENOGENETIC ACTIVATION

2006 ◽  
Vol 18 (2) ◽  
pp. 250
Author(s):  
M. G. Marques ◽  
A. B. Nascimento ◽  
V. P. Oliveira ◽  
A. R. S. Coutinho ◽  
M. E. O. A. Assumpção ◽  
...  
Keyword(s):  
Culture Medium ◽  
Cell Number ◽  
Control Group ◽  
Cleavage Rate ◽  
Vitro Fertilization ◽  
Electric Pulses ◽  
Cell Numbers ◽  
Blastocyst Rate ◽  
And Control

The present work evaluated the reversible meiosis inhibition effect on the development of swine embryos produced by in vitro fertilization (IVF) or parthenogenetic activation (PA). The efficiency of PZM3 and NCSU23 embryo culture media was also evaluated. Oocytes from ovaries collected at a slaughterhouse were subjected to IVM in two different groups: CHX (cycloheximide 5 µM for 10 h) and control, both with TCM-199 + 3.05 mM glucose + 0.91 mM sodium pyruvate + 10% porcine follicular fluid (pFF) + 0.57 mM cystein + 10 ng epidermal growth factor (EGF)/mL + 10 IU eCG/mL + 10 IU hCG/mL for the initial 22 h. In the remaining period (20 h for CHX and 22 h for control), medium without hormones was utilized. After IVM, oocytes were denuded and fertilized for 6 h (IFV) or the matured oocytes were submitted to activation by electric pulses (PA) (2 DC of 1.5 kV/cm for 30 µs), incubated for 1 h in culture medium with 10 μM of CHX, and again submitted to the same electric pulses for 60 µs. Embryo development was evaluated by cleavage rate on Day 3 and blastocyst rate and blastocyst cell number on Day 7 of culture. Cleavage and blastocyst rates were analyzed by the equality-of-two-ratios test and cell number by the Kruskal-Wallis and Mann-Whitney tests (P < 0.05). In relation to IVF, the PZM3 medium was more efficient than NCSU23 for cleavage rate in the CHX group (PZM3: 68.4%, NCSU23: 44.4%) and had a better blastocyst rate in the control group (PZM3: 13.4%, NCSU23: 5.6%). With reference to PA, NCSU23 presented better cleavage and blastocyst rates than PZM3 in the CHX group (NCSU23: 89.5%, PZM3: 78.5% and NCSU23: 20.4%, PZM3: 13.0%, respectively). In the control group, only the NCSU23 blastocyst rate was higher than that for PZM3 (NCSU23: 22.5%, PZM3: 10.8%). No culture medium effect on cell number mean of IVF and PA blastocysts was observed. Maturation block improved cleavage rates in IVF groups cultured with PZM3 (68.4% and 50.6%, respectively, for CHX and control) and in PA groups cultured with NCSU23 (89.5% and 80.3%, respectively, for CHX and control), but no improvement of blastocyst rates in both groups (IVF and PA) was verified. Table 1 below shows that maturation block decreased the IVF and increased the PA blastocyst cell numbers. As older oocytes are more effectively activated, oocytes blocked with CHX achieved the maturation stage faster than the control group, therefore resulting in high-quality PA blastocysts. In conclusion, PZM3 was more efficient for IVF embryo production in contrast to NCSU23, whereas NCSU23 can be indicated for PA embryo production. Moreover, maturation blockage with CHX influenced blastocyst cell number, decreasing in IVF embryos and increasing in PA embryos. Table 1. Mean (±SD) of blastocyst cell numbers for IVF or PA groups after in vitro maturation without (control) or with cycloheximide (CHX) and cultured in NCSU23 or PZM3 medium This work was supported by FAPESP 02/10747–1.

Effect of serum on the in vitro maturation of canine oocytes

1999 ◽  
Vol 11 (8) ◽  
pp. 387 ◽  
Author(s):  
Takeshige Otoi ◽  
Maya Fujii ◽  
Aya Ooka ◽  
Masaki Tanaka ◽  
Tatsuyuki Suzuki
Keyword(s):  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
The Other ◽  
Serum Supplement ◽  
Meiotic Competence ◽  
Positive Effect ◽  
Serum Type

The meiotic competence of canine oocytes cultured for 72 h in medium supplemented with three different concentrations (5, 10 and 20%) of anoestrous, oestrous or metoestrous bitch serum, or with 0.3% bovine serum albumin (BSA), was examined. The oestrous serum supplement had a positive effect on the resumption of meiosis, compared with the other supplements (P<0.05). The number of oocytes that reached metaphase I (MI) and metaphase II (MII) was significantly higher (P<0.05) with the oestrous serum supplement than with the anoestrous serum supplement. There were no significant differences among the three different concentrations in each serum type with respect to the proportion of oocytes that completed meiosis (MI to MII). The number of oocytes that resumed meiosis in the 10% oestrous serum supplement was significantly higher (P<0.05) than those of each concentration of the anoestrous and metoestrous serum supplements, and of the 0.3% BSA supplement. Moreover, a higher number of oocytes reached MII in the presence of the 10% oestrous serum supplement than with the 10% anoestrous serum supplement. These results suggest that supplementation of the culture medium with 10% oestrous serum is the optimal treatment for in vitro maturation of canine oocytes.

scholarly journals Application of nanomaterial for maturation in vitro of cattle embryos

2021 ◽  
Vol 28 ◽  
pp. 112-116
Author(s):  
P. A. Trotskyi ◽  
O. V. Shcherbak ◽  
S. I. Kovtun
Keyword(s):  
In Vitro Maturation ◽  
Cellular Level ◽  
Control Group ◽  
Vitro Fertilization ◽  
Fragmentation Index ◽  
Maturation In Vitro ◽  
Conservation Of Genetic Resources ◽  
Group B ◽  
And Control

Aim. To evaluate the effectiveness of the use of nanomaterial in the environment for the further development of in vitro embryos derived from frozen-thawed oocytes in the system of conservation of genetic resources of animals at the cellular level. Methods. Biotechnological, cryobiological, morphological, cytogenetic, and statistical methods, as well as methods of statistical data processing were used in the research. Results. Oocyte-cumulus complexes (OCC) of cows were divided into four groups: three experimental, in which the maturation was performed in a medium containing 0.1, 0.01 and 0.001% UFS/sucrose and control - without the addition of nanobiomaterial. In vitro fertilization of pre-mature frozen-thawed ova of cows and subsequent maturation of embryos in the medium with the addition of UFS/sucrose (0.001%) showed an increase in the number of embryos by 16.7-22.1% compared with the addition of 0.1; 0.01% and 13.1% compared to the control group. It was found that the fragmentation rate of 2-cell cattle embryos decreased from 65.0 to 39.8% with a decrease in the concentration of UFS/sucrose from 0.1 to 0.001%. The most stable indicators of the fragmentation index from 78.4 to 50.0% were observed on the fourth day of embryo cultivation in experimental group B. Conclusions. Reducing the concentration of UFS/sucrose from 0.1 to 0.001% in the composition of the medium for in vitro maturation of cattle embryos leads to an increase of 16.7-22.1% in the number of embryos obtained. Keywords: oocyte-cumulus complex, cryopreservation, nanomaterial, in vitro maturation, embryo.

134 TRANSCRIPTOME PROFILE OF BOVINE BLASTOCYSTS DERIVED FROM ALTERNATIVE IN VIVO AND IN VITRO CULTURE CONDITIONS AT SPECIFIC PHASES OF EARLY EMBRYONIC DEVELOPMENT

2012 ◽  
Vol 24 (1) ◽  
pp. 179 ◽  
Author(s):  
A. Gad ◽  
U. Besenfelder ◽  
V. Havlicek ◽  
M. Hölker ◽  
M. U. Cinar ◽  
...  
Keyword(s):  
Embryonic Development ◽  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Expression Patterns ◽  
Culture Conditions ◽  
Control Group ◽  
Transcriptome Profile ◽  
Vitro Fertilization

An understanding of gene expression patterns due to altered environmental conditions during different time points of the pre-implantation period would improve our knowledge on regulation of embryonic development and improve success of embryo culture. The aim of this study was to examine the effect of alternative in vivo and in vitro culture conditions at specific phases of early embryonic development on transcriptome profile of bovine blastocysts. Using nonsurgical endoscopic oviducal transfer technology, 5 different blastocyst groups were produced. The first 2 groups were matured in vitro and then either transferred after maturation or after in vitro fertilization to synchronized recipients. The other 3 groups were matured, fertilized and cultured in vitro until 4-cell, 16-cell and morula stage before transfer. Blastocysts from each group were collected by uterine flushing at Day 7 and pooled in groups of 10. Complete in vitro (IVP) and in vivo blastocysts were produced and used as controls. A unique custom microarray (Agilent) containing 42 242 oligo probes (60-mers) was used over 6 replicates of each group vs the in vivo control group to examine the transcriptome profile of blastocysts. Compared with the in vivo control group, clear dramatic shifts were found in the number of differentially expressed genes (DEG, fold change ≥2) at 2 different time points. The first shift occurred for blastocyst groups that were transferred after in vitro fertilization and before embryonic genome activation (EGA). The second shift occurred for blastocyst groups that were transferred after EGA, as well as for the IVP group. Ontological classification of DEG showed that the more time spent under in vitro conditions, the higher the percentage of DEG involved in cell death and apoptotic processes. Moreover, lipid metabolism was the most significant process affected in the blastocysts transferred after in vitro maturation and blastocysts transferred at 16-cell stage. Most DEG involved in this process were down-regulated. Pathway analysis revealed that signalling pathways were the dominant pathways in all groups except the group that was transferred after in vitro maturation. That group showed significant down-regulation for genes involved in retinoic acid receptors activation pathways. These results showed that fertilization and EGA were the most critical developmental stages influenced by in vitro culture conditions and subsequently affect blastocyst quality, as measured in terms of gene expression patterns. Moreover, we identified molecular mechanisms and pathways that were influenced by altered culture conditions. These findings will enable the examination of strategies for modifying in vitro culture conditions at critical stages that will allow more efficient production of developmentally competent blastocysts.

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.

scholarly journals Solidifying agents and activated charcoal for in vitro culture of Solanum sessiliflorum

2017 ◽  
Vol 52 (11) ◽  
pp. 1123-1126 ◽  
Author(s):  
Filipe Almendagna Rodrigues ◽  
Renata Alves Lara Silva Rezende ◽  
Moacir Pasqual ◽  
Maria Teresa Gomes Lopes
Keyword(s):  
In Vitro Culture ◽  
Activated Charcoal ◽  
Root Length ◽  
Culture Medium ◽  
In Vitro Cultivation ◽  
Fresh Matter ◽  
Number Of Leaves

Abstract: The objective of this work was to evaluate the effects of the solidifying agents agar and phytagel and of activated charcoal on the in vitro cultivation of two maná cubiu (Solanum sessiliflorum) varieties: Thaís and Santa Luzia. The phytotechnical characteristics analyzed included number of leaves, number of roots, shoot and root length, and fresh matter of shoot and root. Regardless of the variety, phytagel was superior to agar as a culture medium. A greater number of leaves and longer shoots were observed in the Santa Luzia variety, in the absence of charcoal. The Thaís variety showed longer shoots and roots in the presence of charcoal.

scholarly journals In vitro culture of Nonea pulla DC. seeds.

2020 ◽  
Author(s):  
E.F. Semenova ◽  
◽  
K.V. Vedernikova ◽  
E.Yu. Schetneva ◽  
◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Subsequent Development ◽  
In Vitro Cultivation ◽  
Germinating Capacity ◽  
The Crimea ◽  
Speed Up ◽  
Seed Hull ◽  
Maximum Development

Nonea pulla DC. is a promising perennial medicinal plant growing in the Crimea. Controlled in vitro cultivation of nonea seeds allows improving the up-to-date techniques of seedlings preparation. The conducted experiments confirmed the low germinating capacity of seeds (5–9 %). To increase this parameter and to speed up the introduction process, we investigated the Nonea pulla in vitro culture. The initial phases of germination were expectedly observed during seeds cultivation. The seed swelling, rupture of pericarp and seed hull, release of germ with cotyledons, dehiscence of cotyledons were detected. Moreover, in some cases, no subsequent development was observed. However, normal germs formed in 60% of cases. Seeds also sprouted without the prior cold stratification. For the following growth, plants required a relatively simple culture medium. The maximum development conditions were reached after 1.0–1.5 months of in vitro cultivation (26±2 °С, illuminance of 2000–3000 lux, 16-hour photoperiod).

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.

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