Centrifugal elutriation of porcine oocytes isolated from the ovaries of newborn piglets.
A two step enzymatic procedure for the isolation of ovarian follicles and oocytes at early stages of development from the ovaries of newborn piglets was devised. The isolated oocytes were then separated from the overwhelming majority of the ovarian somatic cells using a centrifugal elutriation system. Twenty to thirty thousand oocytes were routinely collected after elutriation of the cell suspension derived from a pair of ovaries. In the enriched fraction the ratio of oocyte:somatic cells was between 1:1 and 1:3. The validation of the method as an efficient procedure for the isolation of a large, viable, and highly enriched population of oocytes at early stages of development was provided by uptake studies carried out after each step of the isolation and separation technique and by a comparative analysis of the pattern of structural proteins of the enriched fraction and of fully grown porcine oocytes. The results confirmed that the isolated cells were actually oocytes at early stages of development and that they were viable throughout the entire procedure.
Lazzari
G
G
Department of Molecular Embryology, AFRC Institute of Animal Physiology and Genetics Research, Babraham, Cambridge, United Kingdom.
Galli
C
C
Moor
R M
RM
D000818:Animals; D000831:Animals, Newborn; D002469:Cell Separation; D002498:Centrifugation; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008715:Methionine / Q000378:metabolism; D010053:Ovary / Q000166:cytology*; D010063:Ovum / Q000166:cytology* / Q000378:metabolism; D013552:Swine
D008715:Methionine