0.857
IF5
0.900
IF
Q3
JCR
0.92
CiteScore
0.405
SJR
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SJR
20
MNiSW
165.24
ICV
ORIGINAL PAPER
 
CC-BY 4.0
 
 

Vitrification of GV-porcine oocytes by 1-, 2-, and 7-step saturation in ethylene glycol

V. Isachenko 1, 2,  
E. Isachenko 3,  
 
1
Department of Animal Reproduction, Institute for Animal Science, RO.Kulinichi, 312120 Kharkov, Ukraine
2
Department of Animal Biology, Valencia University, Dr. Moliner 50, 46100 Burjassot, Catalonia
3
Department of Human Reproduction, Institute for Problems of Cryobiology and Cryomedicine, Perejaslavs'ka 23, Kharkov, Ukraine
J. Anim. Feed Sci. 1998;7(2):219–230
Publish date: 1998-03-25
KEYWORDS:
ABSTRACT:
Three experiments were conducted to investigate the effect of 1 -, 2- and 7-step saturation with cryoprotectant on vitrification of germinal vesicle porcine oocytes to M-II stage. In Experiment I non-cooled cumulus-oocyte complexes (COCs) were equilibrated in ethylene glycol (EG) using a single-step or step-wise protocol, EG was removed by single-step or step-wise procedure with all manipulations being performed at 42°C. In Experiment II non-cooled COCs were equilibrated in ethylene glycol (EG) using the single-step or step-wise protocol with further EG removal by the single-step or step-wise procedure and pretreatment with Cytochalasin B. In experiment III, performed taking into account the results of Experiments I and II, a total of 177 COCs were divided into three treatment and one control groups. All treatment COCs were vitrified after pretreatment with Cytochalasin B (5 µg/ml at 38.5°C for 10-15 min), direct plunging into liquid nitrogen, thawing for 5 sec in a waterbath at 50°C, removal of the cryoprotectant by exposure to 1M, 0.75M, 0.5M, 0.25M for 4, 2, 2, and 2 min respectively at 40-42°C. Group 1 COCs (n = 52) were cryopreserved after exposition to 40% EG for 4 min. Group 2 COCs (n = 47) were exposed to 25% EG for 5 min and then to 40% EG for 30 sec prior to vitrification. Group 3 COCs (n = 48) were equilibrated over 13 min using a series of EG concentrations 5, 10, 15, 20,25, 30, and 40% EG for 5, 2, 2, 2, 1, 0.5, and 0.5 min, respectively. Group 4 COCs (n = 30) were control oocytes (not exposed to EG and not frozen). After thawing and passage through the sucrose series (treatment) or without treatment (control) COCs were cultured for 48 h in TCM-199 with 10% heat-inactivated follicular fluid and 1 µg/ml FSH in 5% CO 2 at 39°C. The developmental capacity (progression to M-II stage) as assessed by aceto-orcein staining was 0, 4.2, 29.2, and 86.6%, respectively (P1-2 < 0.01; P 2-3, 3-4, 2-4 <0.001).
 
CITATIONS (1):
1. Ultrastructure of Centrifuged Bovine Oocyte-Cumulus Complexes after Pre-treatment with Cytoskeletal Relaxant
V. Isachenko, J. L. Alabart, H. W. Michelmann, N. Bezugly, E. Isachenko, C. Soler, F. Nawroth
Anatomia, Histologia, Embryologia
ISSN:1230-1388