[Zbrafish] in vitro maturation

[Roland Dosch]

Hi...

we culture our oocytes in  60% Leibovitz-L15 medium pH 7.5 with 10mM  
Tris pH 7.5 + 100µg/ml Gentamycin at 26ûC. You have to be careful,  
when you single out oocytes from the ovary. After the surgery leave  
them for an hour in medium. The boken oocytes are indicated by  
leaking ooplasm and should be removed.  These broken oocytes die and  
then look similar to immature oocytes. You don't need to pretreat  
them with hCG.  We add  to a final conc. 1µg/ml DHProgesterone (5mg/ 
ml stock in EtOH=1:5000). After 3 hrs you should see the first clear/ 
mature oocytes.
Good luck,

Roland

______________________________________
Roland DOSCH
Departement de Zoologie et biologie animale
Sciences III
Universite de Geneve
30, quai Ernest-Ansermet
CH-1211 Geneve 4
Switzerland

http://www.biani.unige.ch/dosch

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