Extrusion and removal of lipid from the cytoplasm of porcine oocytes at the germinal vesicle stage: Centrifugation under hypertonic conditions influences vitrification

In the present study, we examined a novel lipid removal method, centrifugation in solutions made hypertonic by adding 0.27 M sugar. This allowed the lipid to be extruded and removed without the loss of active mitochondria or extra cytoplasm. The type of sugar influenced the proportion of oocytes tha...

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Bibliographic Details
Published in:Cryobiology 2005-04, Vol.50 (2), p.216-222
Main Authors: Hara, Kenshiro, Abe, Yasuyuki, Kumada, Nobuyuki, Aono, Nobuya, Kobayashi, Jin, Matsumoto, Hiromichi, Sasada, Hiroshi, Sato, Eimei
Format: Article
Language:English
Subjects:
Animals
Cell Fractionation
Centrifugation
Cryopreservation - methods
Cryopreservation - veterinary
Cytoplasm - chemistry
Cytoplasm - ultrastructure
Female
Glucose Solution, Hypertonic - pharmacology
GV oocytes
Hypertonic condition
Lipid droplets
Lipids - isolation & purification
Micromanipulation
Mitochondria - ultrastructure
Oocytes - chemistry
Oocytes - drug effects
Oocytes - ultrastructure
Pig
Swine
Vitrification
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Summary:In the present study, we examined a novel lipid removal method, centrifugation in solutions made hypertonic by adding 0.27 M sugar. This allowed the lipid to be extruded and removed without the loss of active mitochondria or extra cytoplasm. The type of sugar influenced the proportion of oocytes that could be stratified by centrifugation. Glucose induced the highest extrusion rate of lipid droplets. After vitrification the rates of survival, germinal vesicle breakdown and metaphase II were 30, 26, and 7%, respectively, for lipid-removed GV oocytes; this was significantly higher ( P < 0.05) than for corresponding vitrified lipid-intact oocytes (2, 0, and 0%, respectively). These results indicated that this method is useful to remove whole lipid droplets without losing mitochondria and improves cryotolerance of porcine GV oocytes.
ISSN:0011-2240
1090-2392
DOI:10.1016/j.cryobiol.2005.01.003