scholarly journals Selection of In Vitro-Matured Porcine Oocytes Based on Localization Patterns of Lipid Droplets to Evaluate Developmental Competence

2013 ◽  
Vol 59 (4) ◽  
pp. 405-408 ◽  
Author(s):  
Kou HIRAGA ◽  
Yumi HOSHINO ◽  
Kentaro TANEMURA ◽  
Eimei SATO
Keyword(s):  
Lipid Droplets ◽  
Developmental Competence ◽  
Selection Of

The analysis of chromatin organisation allows selection of mouse antral oocytes competent for development to blastocyst

Zygote ◽  
2002 ◽  
Vol 10 (1) ◽  
pp. 73-78 ◽  
Author(s):  
Maurizio Zuccotti ◽  
Rubén H. Ponce ◽  
Michele Boiani ◽  
Stefano Guizzardi ◽  
Paolo Govoni ◽  
...  
Keyword(s):  
Developmental Competence ◽  
Farm Animals ◽  
Cell Stage ◽  
Developmental Potential ◽  
Different Types ◽  
Chromatin Organisation ◽  
Selection For ◽  
Gamete Selection ◽  
Selection Of

Mouse antral oocytes can be classified in two different types termed SN or NSN oocytes, depending on the presence or absence, respectively, of a ring of Hoechst 33342-positive chromatin surrounding the nucleolus. The aim of the present study was to test the developmental competence to blastocyst of the two types of oocytes. Here we show that following isolation, classification and culture of cumulus-free antral oocytes, 14.7% and 74.5% of NSN and SN oocytes, respectively, reached the metaphase II stage. When fertilised and further cultured none of the metaphase II NSN oocytes developed beyond the 2-cell stage whilst 47.4% of the metaphase II SN oocytes reached the 4-cell stage and 18.4% developed to blastocyst. The findings reported in this paper may contribute to improved procedures of female gamete selection for in vitro fertilisation of humans and farm animals. Furthermore, the selection of oocytes with better developmental potential may be of interest for studies on nuclear/cytoplasm interaction, particularly in nuclear-transfer experiments.

Selection of pre- versus postpubertal pig oocytes for parthenogenetic activation and somatic cell nuclear transfer

2015 ◽  
Vol 27 (3) ◽  
pp. 544 ◽  
Author(s):  
H. S. Pedersen ◽  
Y. Liu ◽  
R. Li ◽  
S. Purup ◽  
P. Løvendahl ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Cell Number ◽  
Developmental Competence ◽  
In Vitro Production ◽  
Oocyte Growth ◽  
Parthenogenetic Activation ◽  
Selection Of

Pig oocytes have been used increasingly for in vitro production techniques in recent years. The slaughterhouse-derived oocytes that are often used are mostly of prepubertal origin. The aims of the present study were to compare the developmental competence between pre- and postpubertal pig oocytes, and to develop a simple and practical method for the selection of prepubertal pig oocytes for parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) based on oocyte morphology after IVM and oocyte inside zona pellucida (ZP) diameter (‘small’ ≤110 µm; ‘medium’ >110 µm; ‘large’ ≥120 µm). Meiotic competence and blastocyst rates after PA and SCNT of prepubertal oocytes increased with oocyte size, with the large prepubertal oocytes reaching a level similar to postpubertal oocytes after SCNT. Blastocyst cell number was not related to oocyte inside ZP diameter and oocyte donor to the same extent as blastocyst rate. Very low blastocyst rates were obtained after PA of morphologically bad pre- and postpubertal oocytes. In conclusion, measurement of inside ZP diameter combined with morphological selection is useful to remove incompetent oocytes. Further studies are needed to clarify the relative importance of cytoplasmic volume and stage in oocyte growth phase.

scholarly journals Lipid content, mitochondrial activity and early embryo development in oocyte collected from crossbred cows (bos taurus indicus)

2018 ◽  
Vol 25 (1) ◽  
pp. 120-131
Author(s):  
Yoeli Mendez ◽  
Nohely Parra ◽  
Francisco Baez ◽  
Robert Valeris ◽  
Patricia Villamediana
Keyword(s):  
Lipid Content ◽  
Lipid Droplets ◽  
Oocyte Quality ◽  
Early Embryo ◽  
Developmental Competence ◽  
Mitochondrial Activity ◽  
Vitro Fertilization ◽  
Immature Oocytes ◽  
Crossbred Cows

The objective of this research was to evaluate the effect of phenotypic predominance on lipid content, mitochondrial activity and early developmental competence as indicators of oocyte quality. Cumulus-oocyte complexes (COCs) were recovered through follicular aspiration, and underwent in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC) of presumptive zygotes. Lipid content and mitochondrial activity in immature and IVM oocytes were determined. A maturation rate of 80.6% and 69.3% was found for oocytes predominantly B. indicus and predominantly B. taurus, respectively. Total fertilization rate was 27.6%; 26.1% for predominantly B. indicus oocytes and 29% for predominantly B. taurus oocytes. A total of 55.5% and 57.5% of cleaved embryos after 48 and 72 h post-insemination (hpi) in predominantly B. indicus group were observed, respectively. As for the predominantly B. taurus group, 48.6% and 60.4% of cleaved embryos were found after 48 and 72 hpi, respectively. In both groups, immature oocytes showed a greater amount of small lipidic droplets (p <0.0001); IVM decreased the number of small lipid droplets (p < 0.0001) and increased the number of medium and large lipid droplets (p < 0.0001). Predominantly B. indicus oocytes had a greater number of small and medium-sized lipid droplets, while there were no significant differences in large lipid droplets. IVM oocytes had higher mitochondrial activity than immature oocytes group (p < 0.05) without any effect of phenotypic predominance on this parameter. Assessment of lipid content was not a predictive factor of oocyte quality in crossbred cows.

Abnormal changes in mitochondria, lipid droplets, ATP and glutathione content, and Ca2+ release after electro-activation contribute to poor developmental competence of porcine oocyte during in vitro ageing

2009 ◽  
Vol 21 (2) ◽  
pp. 323 ◽  
Author(s):  
Ze-Dong Hao ◽  
Shen Liu ◽  
Yi Wu ◽  
Peng-Cheng Wan ◽  
Mao-Sheng Cui ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Developmental Competence ◽  
Atp Content ◽  
Culture Time ◽  
Transmission Electron ◽  
Porcine Oocyte ◽  
Mitochondrial Distribution ◽  
In Vitro Ageing ◽  
Time Required

The present study aims to investigate major changes in porcine oocytes during ageing in vitro. After the oocytes were cultured for 44, 56, 68 and 80 h, changes to porcine oocytes in ultrastructure, mitochondrial distribution, glutathione (GSH) and ATP content, Ca2+ release patterns and developmental competence after electro-activation were observed. Mitochondria were evenly distributed in oocytes at 44 h, aggregated in clusters or in peripheral cytoplasm at 68 h and dimly dispersed throughout ooplasm at 80 h. Mitochondrial shape during ageing was also observed by transmission electron microscopy (TEM) at the same time intervals. Most mitochondria were spherical at 44 h, and became elongated when the culture time was extended to 68 h and 80 h. Moreover, mitochondrial clustering became increasingly loose from 56 h. Lipid droplets in oocytes appeared prominent and electron-dense at 44 h, but electron density was lost at 56 h. Lipid droplets were solidified as of 68 h. There was an age-dependent decrease in ATP content per oocyte. Glutathione content per oocyte decreased significantly and remained lower after 56 h. Amplitudes of [Ca2+] rise decreased dramatically following 56 h, and the time required for [Ca2+] to plateau became shorter after electro-activation with prolonged culture time. Cleavage and blastocyst rates of aged oocytes progressively decreased, while the fragmentation rate gradually increased after electro-activation. It is concluded that abnormal changes in mitochondria, lipid droplets, Ca2+ release after electro-activation, and ATP and GSH content in oocytes during ageing may result in poor developmental competence of parthenotes.

scholarly journals In vitro Production of Porcine Embryos: Current Status and Possibilities – A Review

2020 ◽  
Vol 20 (3) ◽  
pp. 775-796
Author(s):  
Katarzyna Poniedziałek-Kempny
Keyword(s):  
Oocyte Maturation ◽  
Culture Media ◽  
Medium Composition ◽  
Developmental Competence ◽  
Current Status ◽  
In Vitro Production ◽  
Maturation Medium ◽  
Vitro Production ◽  
Selection Of

AbstractThis paper presents the current possibilities, state of knowledge and prospects of in vitro production (IVP) of pig embryos, which consists of in vitro oocyte maturation, in vitro fertilization and in vitro embryo culture. In pigs, oocyte maturation is one of the most important stages in the embryo IVP process. It determines the oocyte’s fertilization ability as well as its embryonic development. Through many research studies of the proper selection of oocytes and appropriate maturation medium composition (especially the addition of various supplements), the in vitro maturation of pig oocytes has been significantly improved. Recent studies have demonstrated that modifications of the diluents and in vitro fertilization media can reduce polyspermy. Furthermore, several adjustments of the porcine culture media with the addition of some supplements have enhanced the embryo quality and developmental competence. These updates show the progress of IVP in pigs that has been achieved; however, many problems remain unsolved.

327 UNSATURATED AND SATURATED NON-ESTERIFIED FATTY ACIDS DIFFERENTLY AFFECT LIPID STORAGE AND DEVELOPMENTAL COMPETENCE OF BOVINE OOCYTES

2010 ◽  
Vol 22 (1) ◽  
pp. 319
Author(s):  
H. Aardema ◽  
P. Vos ◽  
H. Knijn ◽  
B. Roelen ◽  
B. Gadella
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Palmitic Acid ◽  
Lipid Droplet ◽  
Lipid Droplets ◽  
Post Partum ◽  
Developmental Competence ◽  
Negative Effects ◽  
Esterified Fatty Acids

Fertility in high-producing dairy cows has declined over the last decades. An increased serum and follicular fluid concentration of non-esterified fatty acids (NEFAs), due to body fat mobilization in the early post partum period, has been postulated as a cause for this fertility decline. NEFA concentrations and composition may change in the environment of the oocyte and thus might affect the storage depots of esterified NEFAs in the oocyte. We exposed COCs to unsaturated (oleic acid) or saturated (palmitic acid) NEFAs during maturation and subsequently examined lipid droplets and developmental competence of the oocytes. COCs from 3-8 mm follicles of slaughterhouse ovaries were cultured in control maturation medium (TCM-199) and medium containing 100, 250, or 500 μM oleic and/or palmitic acid (10 mM fatty acid was bound to 10% BSA fatty acid free). These concentrations were based on in vivo measured NEFA concentrations in follicular fluid in the early post partum period (Leroy et al. 2005 Reproduction 130, 485-495). After 23 h of maturation, COCs were fertilized (450 per group) and cultured till the blastocyst stage, or fixed (80 per group) for lipid droplet staining with C1-BODIPY® 500/510 C12. Confocal microscopy was performed to determine lipid droplet size in (im(mean) and the number of lipid droplets per oocyte. Lipid droplet number and the log of size were analyzed using analysis of variances with condition as fixed factor. Variation was described as the standard error of the mean. Similar concentrations of palmitic or oleic acid had an opposite effect on the size of lipid droplets in oocytes. The number of lipid droplets dramatically decreased in oocytes exposed to 500 μM palmitic acid (178 ± 20), whereas the number increased after exposure to 500 μM oleic acid (554 ± 15). The number of lipid droplets of oocytes exposed to a combination of 250 μM palmitic acid and 250 μM oleic acid (421 ± 23) was comparable with the control and lower oleic and palmitic acid concentrations. Exposure of COCs to palmitic acid during maturation resulted in reduced blastocyst development in a dose-dependent manner (from 18 ± 1.4%, 13 ± 2.4% to 2.8 ± 1.3% after exposure to 500 μM) when compared to control (20 ± 2.2%) or oocytes exposed to oleic acid (from 23 ± 1.6%, 23 ± 3.3% till 28 ± 3.3%). Negative effects of palmitic acid were counteracted by simultaneous exposure to oleic acid during in vitro oocyte maturation (26 ± 5.5%). We conclude that palmitic acid elicited negative effects on early embryonic development, possibly because it induces a reduction in the number of lipid droplets. These adverse effects can be offset by oleic acid during maturation. Moreover a high oleic acid concentration increased the number and size of lipid droplets of oocytes. The regulatory pathways involved in the noted differences in lipid storage features of in vitro-matured oocytes as well as the adverse effects of palmitic acid on early embryonic development are currently under research.

209 MATURATION KINETICS AFTER HOLDING EQUINE OOCYTES IN EMBRYO HOLDING MEDIUM

2016 ◽  
Vol 28 (2) ◽  
pp. 235
Author(s):  
P. Dini ◽  
O. Bogado ◽  
K. Smits ◽  
A. VanSoom ◽  
P. Daels
Keyword(s):  
In Vitro Maturation ◽  
Polar Body ◽  
Developmental Competence ◽  
Hoechst 33342 ◽  
Immature Oocytes ◽  
Maturation Rate ◽  
Chromatin Configuration ◽  
One Way Anova ◽  
Selection Of

It has been reported that immature, equine oocytes can be maintained in meiotic arrest at 24°C. To evaluate a commercial equine embryo holding medium for storage of equine oocyte at 24°C and to determine the effect of holding on maturation kinetics, cumulus‐oocyte complexes (COC) were recovered from slaughtered mares and placed in Syngro® Embryo Holding Solution at 22–25°C for 18–20 h (OH Group) or placed directly in DMEM-F12-based in vitro maturation (IVM) conditions (D-Mat Group) at 5% CO2 in air at 38.5°C. Maturation rate (metaphase II percentage; MII%) was assessed (presence of polar body under stereomicroscope) after denudation at 22, 24, and 28 h. After assessment, the denuded oocytes that were considered immature were placed back in IVM, reassessed at 24 and 28 h, and MII% was compared with that of oocytes remaining uninterrupted in IVM for 24 and 28 h. One-way ANOVA was used to compare dependent variable in different groups using PROC ANOVA (SAS, version 9.2, SAS Institute, Cary, NC, USA). A random selection of mature oocytes from both groups were fertilised using intracytoplasmic sperm injection (ICSI). A total of 250 injected oocytes were cultured in DMEM-F12 with 10% FCS. Blastocyst rates in OH and D-mat groups were similar (7.1% v. 6.3%). At 22 h, significantly more oocytes reached the MII stage in the OH group than in the D-Mat group, but MII% was similar in both groups at 24 and 28 h (Table 1). Denuded, immature oocytes reached similar maturation rate as the undenuded oocytes in the same group. Our data suggest that oocytes can be held in Syngro® Embryo Holding Solution at 22–25°C for 18–20 h without compromising oocyte developmental competence. Overnight holding of oocytes accelerates maturation with similar maturation rate at 22, 24, and 28 h of IVM in the OH group. Denudation of immature oocytes after 22 h of IVM and returning the denuded oocytes to IVM does not affect the progression of maturation. In subsequent experiment, overnight held oocytes were fixed and stained (Hoechst 33342) and MII% was evaluated after 20, 22, and 28 h of IVM. Chromatin configuration confirmed that stored oocytes reach the MII stage at 22 h. Maturation rates were significantly lower at 20 h, suggesting that 22 h of IVM is required for stored oocytes. Table 1.Maturation rates (% in MII stage) at 22, 24, and 28 h of IVM for equine oocytes held in Syngro® Embryo Holding Medium before IVM (OH) and oocytes placed directly in IVM (D-Mat) Thanks to I. Lemahieu and P. Van Damme. Study was supported by the Special Research Fund at UGent.

scholarly journals Tailoring the gastrointestinal fate of emulsions through interfacial engineering: Impact of lipid type and coating lipid droplets with protein-surfactant mixture

2018 ◽  
Author(s):  
Dayane Dias Cardoso ◽  
Rosiane Lopes da Cunha ◽  
Andresa Gomes
Keyword(s):  
Lipid Droplets ◽  
Surfactant Mixture ◽  
Rational Selection ◽  
Surfactant Mixtures ◽  
Interfacial Engineering ◽  
Oil In Water ◽  
Selection Of

The bioaccessibility of bioactives incorporated in oil-in-water (O/W) emulsions may be modulated by controlling emulsion structure through the rational selection of an oily phase and emulsifier or mixture of emulsifiers. In this context, emulsions were produced with protein-surfactant mixtures and different oils to understand how the nature of ingredients and the interaction between them can affect the structure of emulsions and their behavior during digestion in vitro.

scholarly journals Dielectrophoretic behavior of in vitro-derived bovine metaphase II oocytes and zygotes and its relation to in vitro embryonic developmental competence and mRNA expression pattern

Reproduction ◽  
2007 ◽  
Vol 133 (5) ◽  
pp. 931-946 ◽  
Author(s):  
Salilew-Wondim Dessie ◽  
Franca Rings ◽  
Michael Hölker ◽  
Markus Gilles ◽  
Danyel Jennen ◽  
...  
Keyword(s):  
Polar Body ◽  
Mrna Level ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Non Invasive ◽  
First Polar Body ◽  
Blastocyst Rate ◽  
Selection Of ◽  
Metaphase Ii Oocytes

Selecting developmentally competent oocytes and zygotes based on their morphology is more often influenced by personal judgments and lacks universal standards. Therefore, this experiment was conducted to investigate the rate of development and mRNA level of dielectrophoretically separated oocytes and zygotes to validate dielectrophoresis (DEP) as non-invasive option for selection of oocytes and zygotes. In the first experiment, metaphase II oocytes with (PB+) and without (PB−) first polar body and zygotes were subjected to DEP at 4 MHz and 450 μm electrode distance and classified into fast, very fast, slow, and very slow depending on the time elapsed to reach one of the electrodes in the electric field. Parthenogenetic activation was employed to monitor the embryonic development of dielectrophoretically classified oocytes. The result revealed that at 6 and 7 days of post-activation, the blastocyst rate of very slow dielectrophoretic PB+and PB−oocytes was significantly (P< 0.05) lower than other groups. Similarly, in zygotes, the blastocyst rate at 7 days post-insemination was higher (P< 0.05) in the very fast dielectrophoretic categories when compared with the slow and very slow categories. In the second experiment, mRNA level was analyzed in the very fast and very slow dielectrophoretic PB+oocytes and zygotes respectively using the bovine cDNA microarray. The result showed that 36 and 42 transcripts were differentially regulated between the very fast and very slow dielectrophoretic categories PB+oocytes and zygotes respectively. In conclusion, dielectrophoretically separated oocytes and zygotes showed difference in the rate of blastocyst development accompanied by difference in transcriptional abundances.

Expression levels of FSHR, IGF1R, CYP11al and HSD3β in cumulus cells can predict in vitro developmental competence of bovine oocytes

Zygote ◽  
2020 ◽  
Vol 28 (5) ◽  
pp. 425-431 ◽  
Author(s):  
Atchalalt Khurchabilig ◽  
Akane Sato ◽  
Shiori Ashibe ◽  
Asuka Hara ◽  
Rika Fukumori ◽  
...  
Keyword(s):  
Follicle Stimulating Hormone ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Mrna Levels ◽  
Expression Levels ◽  
Non Invasive ◽  
Production Technologies ◽  
Bovine Oocytes ◽  
Selection Of

SummaryThe efficiency of in vitro embryo production technologies would be improved by the development of suitable non-invasive biomarkers that allow the selection of good quality cumulus–oocyte complexes (COCs). The present study used whole, single oocyte culture to investigate whether the expression levels of follicle-stimulating hormone receptor (FSHR), insulin-like factor 1 receptor (IGF1R) and three steroidogenesis-related enzymes (CYP11al, CYP19al and HSD3β) in cumulus cells reflected the developmental competence of COCs. Cumulus cells were collected from single COCs before maturation culture and relative mRNA levels were assessed using real-time PCR. The analysis indicated that mRNAs for FSHR, IGF1R, CYP11al and HSD3β were present at higher levels in cumulus cells from COCs that failed to form blastocysts compared with cumulus cells from COCs that formed blastocysts. Moreover, FSHR and IGF1R mRNA levels were positively correlated with those of genes for steroidogenesis-related enzymes. In conclusion, poor developmental competence of COCs was related to higher expression of FSHR, IGF1R, CYP11al and HSD3β in cumulus cells, which may indicate the advanced differentiation of cumulus cells into granulosa cells.

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