Effect of melatonin in culture medium on in vitro maturation and DNA integrity of bovine oocytes

2011 ◽  
Vol 21 (1) ◽  
pp. 44-50
Author(s):  
Saadia A. Ali ◽  
Nermeen A. Helmy ◽  
B. R. Abdel-Halim
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Dna Integrity ◽  
Bovine Oocytes

scholarly journals Effect of Granulosa Cells Added to Culture Medium for in Vitro Maturation and Fertilization of Bovine Oocytes.

1991 ◽  
Vol 37 (4) ◽  
pp. 263-271
Author(s):  
Hitoshi MOCHIZUKI ◽  
Yutaka FUKUI ◽  
Kazue OOSAKI ◽  
Hitoshi ONO
Keyword(s):  
Granulosa Cells ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Bovine Oocytes

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.

237 EXPRESSION OF THE GENES HSP 70.1, ZAR-1, AND MATER IN BOVINE OOCYTES SUBMITTED TO PREMATURATION AND/OR IN VITRO MATURATION

2008 ◽  
Vol 20 (1) ◽  
pp. 198
Author(s):  
P. R. Adona ◽  
F. H. Biase ◽  
F. C. Braga ◽  
T. H. C. De Bem ◽  
R. Rochetti ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Software Tool ◽  
Mineral Oil ◽  
Relative Quantification ◽  
Oocyte Competence ◽  
Level Of Significance ◽  
Bovine Oocytes ◽  
Group Part

The present study aimed to assess the transcripts for the proteins that embryos require, histone 2a (H2a-FZ), heat shock 70 kDa protein 1A (HSP 70.1), zygote arrest 1 (ZAR-1), and maternal antigen (MATER), in bovine oocytes submitted to prematuration (PM) culture and/or in vitro maturation (IVM). Follicles (2–6 mm diameter) were aspirated from slaughterhouse-derived ovaries. Oocytes were selected and randomly distributed among treatments. For PM, oocytes were cultured 24 h in TCM-199 medium supplemented with 10 µm butyrolactone I, 0.2 mm pyruvate, and 10 µg mL–1 gentamicin (BGV group). Part of the prematured oocytes were washed and transferred to IVM culture (BMII group). For IVM, oocytes were cultured in TCM-199 supplemented with 10% FCS, 5.0 µg mL–1 LH, 0.5 µg mL–1 FSH, 0.2 mm pyruvate, and 10 µg mL–1 gentamicin for 22 h. As controls one group of oocytes was collected immediately after aspiration (GV group) and another group was matured in vitro (MII group) without undergoing prematuration. All cultures were carried out in 100-µL droplets of culture medium under mineral oil at 38.5�C and 5% CO2 in air. Oocytes from each treatment were denuded and frozen at 80�C (3 pools of 200 oocytes) in PBS with 0.1% polyvinyl alcohol (PVA) and 1 UI µL–1 RNase inhibitor. RNA was extracted using the RNeasy Protect Kit (Qiagen, S�o Paulo, Brazil) according to the manufacturer's instructions. The extracted RNA was used for reverse transcription by the enzyme Improm-II Reverse Transcriptase (Promega, Madison, WI, USA) according to the manufacturer's instructions. cDNA was produced in a thermocycler for 60 min at 42�C, followed by warming to 70�C for 15 min and cooling to 4�C for freezing at –20�C. Relative quantification of the transcripts for the genes H2a-FZ, HSP 70.1, ZAR-1, and MATER was performed by real-time PCR using the SYBR GREEN Kit (Applied Biosystems do Brasil, Sao Paulo, Brazil) according to manufacturer's instructions. Data were analyzed by the REST� software (Relative Expression Software Tool; Pfaffl et al. 2002 Nucl. Acids Res. 30, e36) 2005 BETA V1.9.9; a level of significance of 5% was considered to show differences among transcripts using H2a-FZ to normalize data. No differences were observed (P < 0.05) for the transcripts HSP 70.1, ZAR-1, and MATER, respectively, when comparing GV (1.0, 1.0, and 1.0), MII (0.37 � 0.1, 0.37 � 0.05, and 0.43 � 0.1), and GV with BGV (0.43 � 0.15, 0.54 � 0.2, and 0.33 � 0.25). However, a difference was detected (P < 0.05) between BGV (1.0, 1.0, and 1.0) and BMII (0.17 � 0.1, 0.13 � 0.05, and 0.3 � 0.2), but not between (P > 0.05) MII (1.0, 1.0, and 1.0) and BMII (0.52 � 0.25, 0.5 � 0.1, and 0.6 � 0.25) for transcripts HSP 70.1, ZAR-1, and MATER, respectively. A reduction in transcripts in group BMII may influence oocyte competence, reducing embryo development and/or quality.

Development to the late morula or blastocyst stage following in vitro maturation and fertilization of bovine oocytes

1989 ◽  
Vol 18 (1-3) ◽  
pp. 139-148 ◽  
Author(s):  
Y. Fukui ◽  
M. Urakawa ◽  
C. Sasaki ◽  
N. Chikamatsu ◽  
H. Ono
Keyword(s):  
In Vitro Maturation ◽  
Blastocyst Stage ◽  
Bovine Oocytes
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