Effects of Fructose in a Chemically Defined Maturation Medium on Oocyte Maturation and Parthenogenetic Embryo Development in Pigs

2017 ◽  
Vol 32 (3) ◽  
pp. 139-146
Author(s):  
Hyeji Shin ◽  
Minji Kim ◽  
Joohyeong Lee ◽  
Seung Tae Lee ◽  
Choon-Keun Park ◽  
...  
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
Maturation Medium ◽  
Parthenogenetic Embryo

Oocyte activation procedures and influence of serum on porcine oocyte maturation and subsequent parthenogenetic and nuclear transfer embryo development

Zygote ◽  
2008 ◽  
Vol 16 (4) ◽  
pp. 279-284 ◽  
Author(s):  
E. García-Mengual ◽  
J. Alfonso ◽  
I. Salvador ◽  
C-C. Duque ◽  
M-A. Silvestre
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
Oocyte Activation ◽  
Efficient Procedure ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Porcine Oocyte ◽  
Parthenogenetic Embryo ◽  
Nuclear Transfer Embryo ◽  
Parthenogenetic Embryos

SummaryThe viability of SCNT embryos is poor, with an extremely low cloned piglet production rate. In the present work, we studied the effect of three activation protocols based on ionomycin treatment (5 μM ionomycin for 5 min and incubated in 2 mM 6-DMAP for 3.5 h) or electric stimuli (two square wave electrical DC pulses of 1.2 kV/cm for 30 μs) combined or not with 6-DMAP on parthenogenetic embryo development. Oocytes activated by ionomycin plus 6-DMAP showed lower cleavage (47.2 vs. 78.5–81.5; p < 0.05) and blastocyst rates (11.3 vs. 29.2–32.1; p < 0.05) than those activated by electrical and electrical plus 6-DMAP treatments. Also, we studied the effect of addition of serum to maturation medium (0% vs. 10%) on nuclear maturation and further parthenogenetic and SCNT embryo development. We observed in the parthenogenetic embryos that cleavage rates in the serum-free group were significantly higher than in the serum-supplemented group (81.8 vs. 69.6% respectively; p < 0.05), although these differences were not detected in blastocyst rates or blastocyst nuclei numbers. Regarding SCNT embryos, no significant differences were observed in cleavage or blastocyst rates between different experimental groups of SCNT embryos. In conclusion, electrical pulse followed or not by 6-DMAP was found to be an efficient procedure to artificially activate MII porcine oocytes. Moreover, the addition of serum to oocyte maturation media did not seem to improve parthenogenetic or SCNT porcine embryo development.

150 DEVELOPMENT OF ZONA-FREE AND WITH ZONA PARTHENOGENETIC GOAT EMBRYOS IN DIFFERENT MEDIA

2010 ◽  
Vol 22 (1) ◽  
pp. 233
Author(s):  
M. K. Jena ◽  
D. Malakar ◽  
A. K. De ◽  
S. Garg ◽  
Y. S. Akshey
Keyword(s):  
Embryo Development ◽  
Chemical Activation ◽  
Formation Rate ◽  
Electrical Activation ◽  
Cleavage Rate ◽  
Maturation Medium ◽  
Parthenogenetic Embryo ◽  
Oviductal Fluid ◽  
Phosphate Buffered Saline

The present study was carried out to see the developmental efficiency of zona-free and with zona parthenogenetic goat embryos cultured in Research Vitro Cleave from Cook Australia (RVCL), Embryo Development Media (EDM), modified synthetic oviductal fluid (mSOF), and modified Charles Rosenkrans media (mCR2a). Zona-free embryos were cultured in 4 media, whereas with zona embryos were cultured in 3 media except mCR2a. Ovaries were collected from slaughterhouse and oocytes were isolated by puncturing the follicles in medium containing Dulbecco’s phosphate-buffered saline, 3% BSA, and 50 μg mL-1 gentamicin. Oocytes were matured in maturation medium containing TCM-199 (HEPES modified), 0.05 mg mL-1 Na pyruvate, 0.003 mg mL-1 L-glutamine, 5.5 mg mL-1 glucose, 3 mg mL-1 BSA, 5 μg mL-1 FSH, 10 μg mL-1 LH, 1 μg mL-1 estradiol-17β, 50 μg mL-1 gentamicin, and 10% FBS in 5% CO2 in air at 38.5°C. The COC (15 to 20 oocytes) were placed in 100-μL droplets of maturation medium and incubated in a CO2 incubator (5% CO2 in air) with maximum humidity at 38.5°C for 27 h. Matured oocytes were made cumulus free by treatment with hyaluronidase (0.5 mg mL-1) and zona-free by pronase (2 mg mL-1) in zona-free parthenogenesis. Then the oocytes were activated by 5 μM Ca ionophore for 5 min in a CO2 incubator and then treated with 2 mM 6-DMAP for 4 h. Activation was also done by electrical activation with DC 1.78 kV cm-1, 20 μs, and 2 pulses. Then the zona-free oocytes were kept for in vitro culture in 4 types of media such as RVCL, EDM, mSOF, andm CR2a for 7 days in 5% CO2 in air at 38.5°C. The cleavage rate andmorulae formation were observed in RVCL 40.95%, 13.95%, in EDM 46.92%, 14.75%, in mCR2a 56.66%, 5.88%, and in mSOF 48.23%, 14.63%, respectively. The cleavage rate and morulae formation were also found 55.9%, 14.63% during chemical activation and 32%, 12.5% in electrical activation. Hence, better result was found in chemical activation than electrical activation. For with zona parthenogenesis, the matured oocytes were chemically activated by 5 μM Ca ionophore for 5 min and 2 mM 6-DMAP for 4 h. Then the oocytes were cultured in RVCL, EDM, and mSOF in 100-μL micro-drops media for 7 days. The cleavage, morulae, and early blastocyst production rate were as follows: cleavage rate 75.68%, 72.03%, and 57.11%; morulae 44.61%, 30.29%, and 40.22%; and early blastocyst 17.49%, 11.88%, and 25.01% in RVCL, EDM, and mSOF, respectively. Hatched blastocyst formation rate was 6.75%, 5.48%, and 1.15% in RVCL, EDM, and mSOF, respectively. It could be concluded that zona-free parthenogenetic embryos were produced better in EDM medium and with chemical activation. With zona parthenogenetic embryo development was significantly (P < 0.05) higher in RVCL and EDM media.

Effect of Actin Polymerization Inhibitor During Oocyte Maturation on Parthenogenetic Embryo Development and Ploidy in Capra hircus

2013 ◽  
Vol 51 (11-12) ◽  
pp. 944-953 ◽  
Author(s):  
R. Ranjan ◽  
R. K. Singh ◽  
T. Yasotha ◽  
Manish Kumar ◽  
Gopal Puri ◽  
...  
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
Actin Polymerization ◽  
Capra Hircus ◽  
Polymerization Inhibitor ◽  
Parthenogenetic Embryo

191 SUPPLEMENTATION OF MATURATION MEDIUM WITH FOLLICULAR FLUID, EPIDERMAL GROWTH FACTOR AND NEUREGULIN AFFECTS MITOCHONDRIAL DNA REPLICATION, OOCYTE MATURATION AND EMBRYO DEVELOPMENT IN PIGS

2012 ◽  
Vol 24 (1) ◽  
pp. 208
Author(s):  
J. Mao ◽  
K. M. Whitworth ◽  
L. D. Spate ◽  
E. M. Walters ◽  
J. Zhao ◽  
...  
Keyword(s):  
Growth Factor ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Copy Number ◽  
In Vitro Maturation ◽  
Meiotic Maturation ◽  
Mtdna Copy Number ◽  
Maturation Medium

Mitochondria supply the majority of ATP in a cell. Mitochondrial DNA (mtDNA) copy number in oocytes might be used as a marker of viability and might be a key determinant of pre-implantation embryo development. However, little is known about mtDNA copy number changes during porcine oocyte maturation and its regulation by extracellular growth factors. The objectives of the current study were to determine the effects of supplementation of in vitro maturation medium with porcine follicular fluid (pFF; 0, 10, 20 and 30%), epidermal growth factor (EGF; 10 ng mL–1), neuregulin 1 (NRG; 20 ng mL–1) and NRG + IGF1 (insulin-like growth factor-1; 100 ng mL–1 + NRG, 20 ng mL–1) during in vitro maturation on mtDNA copy number, oocyte meiotic maturation and subsequent embryo development after parthenogenic activation. Follicular fluid used for the pFF supplementation experiment was prepared from medium-sized (3–6 mm in diameter) healthy follicles. Cumulus–oocyte complexes (COCs) were collected from antral follicles (3–6 mm in diameter), cultured in LH- and FSH-containing maturation medium for 22 h at 38.5°C, transferred into basic maturation medium without FSH and LH and cultured for another 22 h. The basic maturation medium was TCM-199 supplemented with 0.1% polyvinylalcohol (w/v), 3.05 mM D-glucose, 0.91 mM sodium pyruvate, 10 μg mL–1 of gentamicin, 0.57 mM cysteine and without or with different growth factors depending on the experimental design. In total, 177 germinal vesicle (GV) oocytes and 3837 MII oocytes were used for this study. All data were analyzed by the general linear model (GLM) procedure of SAS software (V9.2). The mtDNA copy number in oocytes increased (P < 0.05) from GV to MII stage oocytes (MII oocytes from all treatment groups pooled). Supplementation of IVM media with 10% pFF decreased mtDNA copy number (P < 0.05), whereas 20 and 30% pFF had no major effect on mtDNA copy number, resulting in a quadratic correlation between percentage of pFF and mtDNA copy number. There was a negative linear correlation between percentage of pFF and oocyte meiotic maturation, with a higher percentage of pFF inhibiting meiotic maturation (73.2 ± 5.2, 71.9 ± 4.8, 64.1 ± 8.5 and 65.8 ± 6.4% for 0, 10, 20 and 30% pFF groups, respectively). The mtDNA copy numbers in EGF and NRG-treated MII oocytes were significantly higher than those in GV oocytes, whereas the control was not different (EGF, 237 042.6 ± 22 198.2; NRG, 281 293.4 ± 22 893.5; and control, 231 856.8 ± 21 883.5 in MII oocytes vs 192 288.7 ± 21 675.4 in GV oocytes). The EGF, NRG and NRG+IGF1 treatments enhanced oocyte maturation as well. There was no difference in Day-7 blastocyst formation between EGF, NRG+IGF1 and the control, whereas the NRG treatment enhanced blastocyst formation as compared to the control (23.8 ± 2.4 vs 15.1 ± 2.1%; P < 0.05). This study demonstrated that there was an increase in mtDNA copy number during in vitro maturation. The EGF and NRG treatments stimulated mitochondria biogenesis, which may provide new means to increase oocyte quality and enhance embryonic development.

scholarly journals Pengaruh Penambahan Chorionic Gonadotrophin pada Medium Maturasi terhadap Kemampuan Maturasi, Fertilisasi, dan Perkembangan Embrio secara In Vitro Kambing Peranakan Ettawa (The Effect of Chorionic Gonadotrophin Addition Into Maturation Medium on The Abili

2012 ◽  
Vol 34 (1) ◽  
pp. 8
Author(s):  
Nurvina Septi Adifa ◽  
Pudji Astuti ◽  
Diah Tri Widayati
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
Chorionic Gonadotrophin ◽  
Fertilization In Vitro ◽  
Cleavage Rate ◽  
Group I ◽  
Maturation Medium ◽  
Frozen Semen ◽  
Significant Difference

<p>This research was conducted to investigate the effect of chorionic gonadotrophin addition into maturation medium on oocyte maturation, fertilization, and embryo development in vitro of Ettawa crossbred. Oocytes were divided into 3 groups, group I: maturation medium without addition of chorionic gonadotrophin (0), group II: 10 μl/10 ml chorionic gonadotrophin was added into maturation medium (1), group III: 20 μl/10 ml chorionic gonadotrophin was added into maturation medium (2). Oocytes were transferred into 50 μl maturation medium, then covered by mineral oil. Oocyte was incubated at 39oC, 5% CO2, 95% humidity for 24 hours for maturation. Matured oocytes were inseminated with frozen semen–thawed concentration 12.5 x 106/ml. Process of fertilization were carried out on incubator 39oC, 5% CO2, 95% humidity for 5 hours. The fertilized oocytes were transferred into 50 μl drop G–1, then incubated at 39oC, 5% CO2, 95% humidity. Embryo development was monitored every 24 hours. Culture medium was changed every 48 hours. G–2 medium used second day after culture. The variables measured involved oocyte maturation, fertilization, and in vitro cleavage rate. The data were analyzed by chi–square, using SPSS 15.0 program. The result showed no significant difference on the percentage of mature oocytes and fertilization rate were 78.0%, 72.8%, 75.0% and 76.6%, 74.5%, 77.8% respectively. But cleavage rate showed significant difference (P≤0.05) with<br />the values of 40.8%, 11.4%, and 12.2% respectively. Based on the result it could be concluded that chorionic gonadotrophin addition into maturation medium had not increased ettawa crossbred oocytes maturation, fertilization, and in vitro cleavage rate. The best maturation, fertilization, and in vitro cleavage rate were found using maturation medium without any addition of chorionic gonadotrophin.</p><p>(Key words: Does oocyte, Chorionic gonadotrophin, In vitro maturation, In vitro fertilization, In vitro embryo development)<br /><br /></p>

Pioglitazone improves porcine oocyte maturation and subsequent parthenogenetic embryo development in vitro by increasing lipid metabolism

2019 ◽  
Vol 86 (9) ◽  
pp. 1245-1254 ◽  
Author(s):  
Sang‐Gi Jeong ◽  
Seung‐Eun Lee ◽  
Won‐Jae Kim ◽  
Yun‐Gwi Park ◽  
Jae‐Wook Yoon ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Porcine Oocyte ◽  
Parthenogenetic Embryo ◽  
Development In Vitro

Rho-GTPase effector ROCK regulates mouse oocyte maturation and embryo development

2014 ◽  
Author(s):  
Xing Duan ◽  
Zhen-Bo Wang ◽  
Xiang-Shun Cui ◽  
Nam-Hyung Kim ◽  
Shao-Chen Sun
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
Rho Gtpase ◽  
Mouse Oocyte

scholarly journals X-Linked Huwe1 Is Essential for Oocyte Maturation and Preimplantation Embryo Development

iScience ◽  
2020 ◽  
Vol 23 (9) ◽  
pp. 101523
Author(s):  
Alaa A. Eisa ◽  
Scott Bang ◽  
Katherine J. Crawford ◽  
Emily M. Murphy ◽  
William W. Feng ◽  
...  
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
Preimplantation Embryo ◽  
Preimplantation Embryo Development
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