In vitro maturation of a mammalian cumulus oocyte complex

Abstract

The present invention relates to a composition and method for assisted reproductive technology in mammals. In particular, the present invention provides compositions and methods for in vivo maturation of an immature cumulus oocyte complex (COC), thereby enhancing the embryology outcome.

The invention claimed is: 
     
       1. A capacitation medium for in vitro maturation of an immature mammalian cumulus oocyte complex, the capacitation medium comprising 0.1-50 nM from 0.1 nM to 100 nM C-type natriuretic peptide (CNP), estradioland, Follicle Stimulating Hormone (FSH) or an FSH analog, and from 0.1 ng/ml to 10 ng/ml insulin. 
     
     
       2. The capacitation medium according to  claim 1 , wherein said medium also comprises 0.1-10 ng/ml insulin. 
     
     
       3. The capacitation medium according to  claim 1 , wherein said medium further comprises comprising an oocyte-secreted factor or combination of oocyte-secreted factors. 
     
     
       4. The capacitation medium according to  claim 1 , wherein said medium comprises an oocyte-secreted factor selected from the group consisting of GDF-9, BMP-15, FGF-8, and combination thereof. 
     
     
       5. A kit for in vitro maturation of an immature mammalian cumulus oocyte complex, the kit comprising the capacitation medium according to  claim 1 . 
     
     
       6. The capacitation medium according to  claim 1 , comprising from 1 nM to 1000 nM estradiol. 
     
     
       7. The capacitation medium according to  claim 1 , comprising from 0.1 mIU/ml to 10 mIU/ml FSH or FSH analog. 
     
     
       8. The capacitation medium according to  claim 1 , further comprising from 10 ng/ml to 100 ng/ml oocyte-secreted factor. 
     
     
       9. The capacitation medium according to  claim 8 , wherein the oocyte-secreted factor is selected from the group consisting of GDF-9, functional equivalents or fragments of GDF-9, BMP-15, functional equivalents or fragments of BMP-15, FGF-8, functional equivalents or fragments of FGF-8, and combinations thereof. 
     
     
       10. A capacitation medium, comprising:
 from 0.1 nM to 100 nM CNP;   from 1 nM to 1000 nM estradiol;   from 0.1 mIU/ml to 10 mIU/ml FSH or FSH analog;   from 0.1 to 10 ng/ml insulin; and   from 10 ng/ml to 100 ng/ml of an oocyte-secreted factor or combination of oocyte-secreted factors.   
     
     
       11. The capacitation medium according to  claim 10 , comprising from 10 nM to 100 nM CNP. 
     
     
       12. The capacitation medium according to  claim 10 , comprising from 0.1 nM to 50 nM CNP. 
     
     
       13. The capacitation medium according to  claim 10 , comprising from 10 nM to 50 nM CNP. 
     
     
       14. The capacitation medium according to  claim 10 , wherein the oocyte-secreted factor is selected from the group consisting of GDF-9, functional equivalents or fragments of GDF-9, BMP-15, functional equivalents or fragments of BMP-15, FGF-8, functional equivalents or fragments of FGF-8, and combinations thereof. 
     
     
       15. The capacitation medium according to  claim 10 , wherein the oocyte-secreted factor is selected from the group consisting of GDF-9, BMP-15, FGF-8, and combinations thereof. 
     
     
       16. A kit for in vitro maturation of an immature mammalian cumulus oocyte complex, the kit comprising the capacitation medium according to  claim 1 . 
     
     
       17. The kit according to  claim 16 , further comprising:
 a collection medium;   a maturation medium;   non-adherent and/or adherent culture plates; and   instructions for use of the kit.   
     
     
       18. The capacitation medium according to  claim 3 , wherein the oocyte-secreted factor or combination of oocyte-secreted factors is selected from the group consisting of GDF-9, functional equivalents or fragments of GDF-9, BMP-15, functional equivalents or fragments of BMP-15, FGF-8, functional equivalents or fragments of FGF-8, and combinations thereof.