scholarly journals Presence of vascular endothelial growth factor during the first half of IVM improves the meiotic and developmental competence of porcine oocytes from small follicles

2017 ◽  
Vol 29 (10) ◽  
pp. 1902 ◽  
Author(s):  
Tra M. T. Bui ◽  
Khánh X. Nguyễn ◽  
Asako Karata ◽  
Pilar Ferré ◽  
Minh T. Trần ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Formation Rate ◽  
Developmental Competence ◽  
Vascular Endothelial ◽  
Blastocyst Formation ◽  
Parthenogenetic Activation ◽  
Maturation Rate ◽  
Endothelial Growth Factor

The aim of the present study was to investigate the effect of vascular endothelial growth factor (VEGF) on the meiotic and developmental competence of porcine oocytes from small follicles (SF; 0.5–3 mm diameter). When cumulus–oocyte complexes (COCs) from medium-sized follicles (MF; 3–6 mm diameter) and SF were cultured for IVM, the maturation rates were significantly higher for oocytes from MF than SF. Concentrations of VEGF in the medium were significantly higher for COCs cultured from MF than SF. When COCs from SF were exposed to 200 ng mL–1 VEGF during the first 20 h of IVM, the maturation rate improved significantly and was similar to that of oocytes derived from MF. The fertilisability of oocytes was also significantly higher than that of VEGF-free SF controls. Following parthenogenetic activation, the blastocyst formation rate improved significantly when SF COC culture was supplemented with 200 ng mL–1 VEGF, with the rate similar to that of oocytes from MF. The results of the present study indicate that VEGF markedly improves the meiotic and developmental competence of oocytes derived from SF, especially at a concentration of 200 ng mL–1 during the first 20 h of IVM.

99 EFFECT OF VASCULAR ENDOTHELIAL GROWTH FACTOR ON IN VITRO-PRODUCED PORCINE OOCYTES AND THEIR SUBSEQUENT DEVELOPMENT: A PARTHENOGENETIC STUDY

2008 ◽  
Vol 20 (1) ◽  
pp. 130
Author(s):  
D. Biswas ◽  
J. H. Lee ◽  
E. B. Jeung ◽  
E. S. Lee ◽  
S. H. Hyun
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Inner Cell Mass ◽  
Formation Rate ◽  
Cell Mass ◽  
Vascular Endothelial ◽  
Blastocyst Formation ◽  
Group B ◽  
Endothelial Growth Factor

The addition of vascular endothelial growth factor (VEGF) to maturation media has beneficial effects on oocyte maturation and blastocyst formation (Einspanier et al. 2002 Mol. Reprod. Dev. 62, 29–36). The present study was conducted to examine the effect of parthenogenesis on in vitro-matured porcine oocytes with VEGF along with porcine follicular fluid in the maturation media. Porcine ovaries were collected from a local slaughter house in physiological saline. After aspiration, COC were matured in vitro in TCM-199 supplemented with 10 ng mL–1 of epidermal growth factor and (1) Group A: 10% pFF; (2) Group B: 10% pFF and 5 ng mL–1 of VEGF; (3) Group C: 10% polyvinyl alcohol; or (4) Group D: 5 ng mL–1 of VEGF plus 10% polyvinyl alcohol. Fifty COC were cultured for the first 22 h at 390�C in a humidified atmosphere of 5% CO2 in 95% air with 4 IU mL–1 of eCG and 4 IU mL–1 of hCG. They were then transferred to hormone-free medium and cultured for an additional 20 h. After culture, COC were denuded with hyaluronidase, and a proportion were stained with Hoechst 33342 for evaluating the metaphase II stage. The remaining oocytes were subjected to electrical parthenogenesis by using a 1-mm fusion chamber and were activated by applying 2 direct current pulses of 110V for 60 µs. Cleavage and blastocyst formation rate were evaluated under a stereomicroscope at 48 and 168 h after activation, respectively. Blastocyst quality was assessed by differential staining of inner cell mass and trophectoderm cells according to a modified staining procedure (Thouas et al. 2001 Reprod. Biomed. Online 3, 25–29). All data are presented as mean � SD and were analyzed by ANOVA followed by Duncan's multiple range test using SPSS 12.0 (SPSS Inc., Chicago, IL, USA). The maturation rate was significantly higher (P < 0.05) in Groups A and B than Groups C and D (76.1 � 9.6, 78.9 � 6.0 v. 60. � 14.2 and 58.3 � 14.3, respectively). The cleavage rate was significantly higher (P < 0.05) in Groups A and B (73.2 � 1.8 and 64.6 � 1.1, respectively) than Groups C and D (47.9 � 1.8 and 48.3 � 1.7, respectively). The blastocyst formation rate was significantly higher (P < 0.05) in Group B (32.6 � 2.4) compared to other groups. There was no significant difference in blastocyst cell number (inner cell mass or trophectoderm) among these groups. These data indicate that the exogenous VEGF along with pFF in the maturation media helps to increase the blastocyst formation rate in vitro, and it might be due to presence of some ligand/protein kinase in the pFF that plays an important role during the cyclic growth of oocytes.

120 SUPPLEMENTATION OF VASCULAR ENDOTHELIAL GROWTH FACTOR DURING IN VITRO MATURATION OF PORCINE IMMATURE CUMULUS - OOCYTE COMPLEXES AND SUBSEQUENT DEVELOPMENTAL COMPETENCE AFTER PARTHENOGENESIS AND SOMATIC CELL NUCLEAR TRANSFER

2011 ◽  
Vol 23 (1) ◽  
pp. 165 ◽  
Author(s):  
D. Biswas ◽  
Y.-B. Jeon ◽  
G.-H. Kim ◽  
E.-B. Jeung ◽  
S. H. Hyun
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Cell Number ◽  
Total Cell ◽  
Vascular Endothelial ◽  
Blastocyst Formation ◽  
Total Cell Number ◽  
Endothelial Growth Factor

In the present study, pig cumulus–oocyte complexes were cultured in medium supplemented with different concentrations (0, 5, 50, and 500 ng mL–1) of vascular endothelial growth factor (VEGF), and then the maturation and intracellular glutathione (GSH) concentration of oocytes were examined. In addition, the development of oocytes matured with different concentrations of VEGF after parthenogenetic activation (PA) or somatic cell nuclear transfer (SCNT) was observed. Although the maturation rate of oocytes was not affected by VEGF concentrations (81.13 ± 2.61%, 83.93 ± 1.97%, 82.14 ± 4.03%, 75.24 ± 2.68%, respectively), the intracellular GSH concentrations of oocytes matured with 5 and 50 ng mL–1 VEGF were significantly higher (12.68 ± 0.08, 12.33 ± 0.53 pMol/oocyte, respectively) than those of oocytes matured with 0 or 500 ng mL–1 VEGF (10.19 ± 0.66, 10.54 ± 0.54 pMol/oocyte, respectively). The blastocyst formation rates after PA of oocytes matured with 5 and 50 ng mL–1 VEGF were significantly higher (58.99 ± 4.70% and 54.00 ± 1.09%, respectively) than that of oocytes matured with 0 or 500 ng mL–1 VEGF (30.15 ± 4.52%, 34.79 ± 4.01%, respectively). Total cell number of PA blastocyst after oocytes matured with 5 and 50 ng mL–1 VEGF was significantly higher (83.21 ± 4.89, 78.16 ± 6.15, respectively) than that of control and 500 ng mL–1 VEGF (56.91 ± 4.78, 55.93 ± 3.89, respectively). Similarly, the blastocyst formation rate after SCNT of oocytes matured with 5 ng mL–1 VEGF was significantly higher (14.54 ± 1.42%) than that of oocytes matured without VEGF (7.95 ± 1.44%). Total cell number of SCNT blastocyst after oocytes matured with 5 ng mL–1 VEGF was significantly higher (67.83 ± 6.56) than control (48.09 ± 5.36). Fully cumulus cell expansion was significantly higher in the 5 ng mL–1 VEGF treated group (85.37 ± 0.73%) compared with the control (58.89 ± 0.88%). In conclusion, adding 5 ng mL–1 VEGF during IVM improved the developmental potential of PA and SCNT in porcine embryos by increasing the intracellular GSH level. This work was supported by a grant (#20070301034040) from BioGreen 21 program, Rural Development Administration, Republic of Korea.

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