Effects of Granulosa Cell Mitochondria Transfer on the Early Development of Bovine Embryos In Vitro

2007 ◽  
Vol 9 (2) ◽  
pp. 237-246 ◽  
Author(s):  
Song Hua ◽  
Yong Zhang ◽  
Xiang-Chen Li ◽  
Li-Bing Ma ◽  
Jun-Wei Cao ◽  
...  
Keyword(s):  
Granulosa Cell ◽  
Early Development ◽  
Bovine Embryos ◽  
Mitochondria Transfer

scholarly journals 113COMPARISON OF TWO ETHYLENE GLYCOL EQUILIBRATION TREATMENTS FOR THE QUICK FREEZING OF IN VITRO-PRODUCED BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 178
Author(s):  
A.C. Nicácio ◽  
R. Simões ◽  
C. Yamada ◽  
H.V.A. Caetano ◽  
M.R.B. Mello ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Liquid Nitrogen ◽  
Granulosa Cell ◽  
Cell Monolayer ◽  
Slow Freezing ◽  
Bovine Embryos ◽  
Treatment Groups ◽  
Quick Freezing ◽  
Hatching Rates

The aim of this study was to compare two ethylene glycol (EG) equilibration procedures for the quick freezing of in vitro-produced bovine embryos. Cumulus-oocyte complexes (COCs) were collected from slaughterhouse ovaries. COCs were matured in TCM199 containing 10% of bovine fetal serum, LH, FSH and E2, and fertilized. Presumptive zygotes were co-cultured in TCM199 with a granulosa cell monolayer, at 39°C in humidified atmosphere of 5% CO2 in air. Grade 1, expanded blastocysts (n=761) were selected 7 and 9 days after insemination and randomly distributed to one of eight treatment groups. In Equilibration Procedure 1, embryos were exposed to 10% EG for 5 min, and then to 17%, 22% or 28% EG for 60s (respectively referred to as EG 17, EG 22 and EG 28). In Equilibration Procedure 2, embryos were exposed to the same EG solutions as in Equilibration Procedure 1, but the period of exposure was 10min to 10% EG and 30 s to EG 17, EG 22 and EG 28. In Equilibration Procedure 3 (slow-freezing controls), embryos were exposed to 10% EG for either 5 or 10min and then cryopreserved by slow-freezing method at 1.2°C/min. In all treatment groups, EG solutions were prepared in PBS+0.2% BSA, and embryos were exposed to EG solutions at 22°C. Embryos were loaded into 0.25mL straws and heat-sealed. Straws were cooled in liquid nitrogen vapor for 2min, and then plunged into and stored in liquid nitrogen. Straws were thawed in room temperature air for 10s, and then in 25°C water for 20s. Thawed embryos were diluted by transferring them into 0.5ml of PBS+0.2% BSA+0.3M sucrose for 3min, and then 0.5mL of PBS+0.2% BSA for 3min. Embryos were co-cultured on granulosa cell monolayer in TCM199 and evaluated after 24h for blastocyst re-expansion (EXP), and again at 48, 72 and 96h for hatching (HAT). A total of 724 in vitro-produced bovine blastocysts were used as controls to determine hatching rates. The results are presented in the table. Embryos exposed to 10% EG for 10min (Equilibration Procedure 1) yielded significantly higher rates of blastocyst re-expansion and hatching when compared to embryos exposed for 5min (Equilibration Procedure 2, P<0.05). These results suggest that quick freezing of in vitro-derived bovine embryos may be an alternative to vitrification; however, additional studies are needed to optimize cryopreservation protocols and increase post-thaw survival. This project was supported by FAPESP (01/11266-4) Table 1 Effect of equilibration procedure on in vitro re-expansion and hatching rates of embryos cryopreserved by slow and quick freezing methods

scholarly journals 122COMPARISON OF TWO CRYOPROTECTANT DILUTION TREATMENTS FOR QUICK FROZEN IN VITRO-PRODUCED BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 183
Author(s):  
J.A. Visintin ◽  
A.C. Nicácio ◽  
C. Yamada ◽  
H.V.C. Amaral ◽  
R. Simões ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Granulosa Cell ◽  
Sucrose Solution ◽  
Cell Monolayer ◽  
Bovine Embryos ◽  
Treatment Groups ◽  
Nitrogen Vapor ◽  
Quick Freezing ◽  
Hatching Rates

The aim of this study was to compare the viability of in vitro-produced bovine embryos following quick freezing in ethylene glycol (EG) and subsequent dilution of EG by either a two- or a three-step procedure. Cumulus-oocyte complexes (COCs) were collected from slaughterhouse ovaries. COCs were matured in TCM199 containing 10% bovine fetal serum, LH, FSH and E2, and fertilized. Presumptive zygotes were co-cultured in TCM199 with a granulosa cell monolayer, at 39°C in humidified atmosphere of 5% CO2 in air. Grade 1, expanded blastocysts (n=544) were selected 7 and 9 days after insemination and randomly distributed to one of three EG equilibration treatment groups. Embryos were exposed to 10% EG for 10min, and then to 17%, 22% or 28% EG for 30s (respectively referred to as EG 17, EG 22 and EG 28). In all treatment groups, EG solutions were prepared in PBS+0.2% BSA, and embryos were exposed to EG solutions at 22°C. Embryos were loaded into 0.25-mL straws which were then heat-sealed. Straws were cooled in liquid nitrogen vapor for 2min, and then plunged and stored in liquid nitrogen. Straws were thawed in room temperature air for 10s, and then in 25°C water for 20s. The thawed embryos of the EG 17, EG 22 and EG 28 groups were randomly assigned to one of two EG dilution procedures. Two-step dilution consisted of transfer of embryos into PBS+0.2% BSA+0.3M sucrose solution for 3min, and then PBS+0.2% BSA for 3min. Three-step dilution consisted of transfer of embryos into PBS+10% EG+0.2% BSA+0.3M sucrose for 3min, PBS+0.2% BSA+0.3M sucrose for 3min, and then PBS+0.2% BSA for 3min. Embryos were co-cultured on a granulosa cell monolayer in TCM199 and evaluated after 24h for blastocyst re-expansion (EXP), and again at 48, 72 and 96h for hatching (HAT). A total of 724 in vitro-produced bovine blastocysts were used as controls to determine hatching rates. The results are presented in the Table. No significant differences were found between the two- and three-step dilution procedures (P>0.05) for in vitro-produced bovine embryos cryopreserved by quick freezing. This project was supported by FAPESP (01/11266-4). Table 1 In vitro re-expansion and hatching rates (%) of rapidly frozen embryos after two- or three-step dilution

scholarly journals Effect of zona pellucida removal on early development of in vitro produced bovine embryos

2013 ◽  
Vol 45 (1) ◽  
pp. 7-15 ◽  
Author(s):  
AE Velásquez ◽  
JR Manríquez ◽  
FO Castro ◽  
Ll Rodríguez-Alvarez
Keyword(s):  
Early Development ◽  
Zona Pellucida ◽  
Bovine Embryos

STUDIES ON THE GROWTH OF EXPERIMENTAL OVARIAN TUMOURS IN TISSUE CULTURE

1959 ◽  
Vol XXXII (I) ◽  
pp. 41-53 ◽  
Author(s):  
Stig Kullander ◽  
Bengt Källén
Keyword(s):  
Tissue Culture ◽  
Granulosa Cell ◽  
In Vitro Study ◽  
Vitro Study ◽  
Ovarian Tumours

ABSTRACT An in vitro study has been made of experimentally produced rat ovarian tumours of different age, paying particular attention to tumour reaction to crystallized steroids. Tumours of two histological structures were found: granulosa cell – luteoma tumours and arrhenoblastoma tumours. Both types grew in vitro and pictures of their cell appearance are given. The former type gave the best growth, and the endocrine studies were restricted to this type. The steroids tested (androsterone, oestrone, progesterone) all had an arresting effect in certain cases. This effect is not an unspecific, toxic one. The different tumours react to different extents, some being completely unaffected.

Early development of bovine embryos depends on the cooperative action between oxidative and endoplasmic reticulum stresses

2014 ◽  
Author(s):  
Seung-Bin Yoon ◽  
Hae-Jun Yang ◽  
Seong-Eun Mun ◽  
Pil-Soo Jeong ◽  
Seon-A Choi ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Early Development ◽  
Bovine Embryos ◽  
Cooperative Action
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