1. Autologous Reimplantation
of Whole Ovine Ovaries
Martin Imhof
Department of Biomedical Sciences
Medical University of Vienna
Landesklinikum Weinviertel
Korneuburg
martin.imhof@meduniwien.ac.at

2. Cryopreservation of Ovarian Tissue
Follicle loss after implantation
Limited follicle pool

3. Cryopreservation of Ovarian Tissue
Connection to vascular system
Immediate perfusion

4. Cryopreservation of Ovarian Tissue
Size of ovary - Cryopreservation
Major surgical procedure

5. Experiment 1 - Whole Porcine Ovary
J Assist Reprod Genet. 2004 Dec;21(12):459-65. Cryopreservation of a whole ovary as a strategy for restoring
ovarian function. Imhof et al.
controlled fresh plunged

6. Experiment 1 - Whole Porcine Ovary
J Assist Reprod Genet. 2004 Dec;21(12):459-65. Cryopreservation of a whole ovary as a strategy for restoring
ovarian function. Imhof et al.
1,5 M DMSO
10% Albumin
RPMI - Medium

7. Freezing Curve
2°C/min to 0°C
0°C for 12 min.
1.6°C/min to -9°C
8 min at -9°C
by 0.4°C/min to
-40°C
to -150°C at the
faster rate of
10°C/min.

8. controlled frozen
Experiment 1 - Whole Porcine Ovary
J Assist Reprod Genet. 2004 Dec;21(12):459-65. Cryopreservation of a whole ovary as a strategy for restoring
ovarian function. Imhof et al.

9. plunged
Experiment 1 - Whole Porcine Ovary
J Assist Reprod Genet. 2004 Dec;21(12):459-65. Cryopreservation of a whole ovary as a strategy for restoring
ovarian function. Imhof et al.

10. Mitochondria
Endoplasm. Reticula
Experiment 1 - Whole Porcine Ovary
J Assist Reprod Genet. 2004 Dec;21(12):459-65. Cryopreservation of a whole ovary as a strategy for restoring
ovarian function. Imhof et al.

11. Experiment 1 - Whole Porcine Ovary

12. Exp. 2 - Whole ovine ovary
Fertil Steril. 2006 Apr;85 Suppl 1:1208-15. Orthotopic microvascular reanastomosis of whole
cryopreserved ovine ovaries resulting in pregnancy and live birth. Imhof et al.

13. Cryopreservation of a Whole Ovary
- Reimplantation of Sheep Ovaries -
step I: - unilateral ovarectomy
- cryopreservation of the ovary
step II: - contralateral explantation
and
- contralateral orthotopic
reimplantation with
microanastomosis of the ovarian vessels
- FSH-, progesterone and ultrasonic follicle screening

16. Ovary
Vessels
Salpinx
Uterus
Reimplantation of a Whole Ovary
- Vessel Preparation -

17. Reimplantation of a Whole Ovary
- Vessel Preparation -

18. Reimplantation of a Whole Ovary
- Vessel Preparation -

19. Reimplantation of a Whole Ovary
- Vessel Preparation -
artery
vein

20. Reimplantation of a Whole Ovary
- Vessel Preparation -

21. J Assist Reprod Genet. 2004 Dec;21(12):459-65. Cryopreservation of a whole ovary as a strategy for restoring
ovarian function. Imhof et al.
1,5 M DMSO
10% Autologous Serum
RPMI - Medium
Reimplantation of a Whole Ovary
- Cryosolution -

22. Slow Freezing
2°C/min to 0°C
0°C for 12 min.
1.6°C/min to -9°C
8 min at -9°C
by 0.4°C/min to
-40°C
to -150°C at the
faster rate of
10°C/min.

23. Reimplantation of a Whole Ovary
- Anastomosis -
operation
microscope
performing
vascular
anastomosis

24. Transplantation of a Whole Ovary
- Anastomosis -
vessel anastomosis

25. Reimplantation of a Whole Ovary
- Perfusion -

26. Reimplantation of a Whole Ovary
- Perfusion -

27. Reimplantation of a Whole Ovary

28. Reimplantation of a Whole Ovary

29. Reimplantation of a Whole Ovary

30. Reimplantation of a Whole Ovary

31. Reimplantation of a Whole Ovary
Reimpl. + 545 Tage

32. Reimplantation of a Whole Ovary

33. small vessels with
healthy endothelial cells
Reimplantation of a Whole Ovary

34. Reimplantation of Ovine Ovaries
- Results -

35. Reimplantation of Sheep Ovaries
- Results -

36. Cryopreservation of Ovarian Slices
advantage: - simplified freezing technique
- easy ex- and implantation
- orthotopic position possible
- implantation repeatable
disadvantage: - absolute and relative reduced
number of follicles

37. Cryopreservation of Ovarian Slices

38. Ovary
Cryopreservation of Ovarian Slices

39. Radiation of Remaining Ovary

40. Radiation of Remaining Ovary

41. Reimplantation
Kontralaterales
Ovar

42. 8 reimplantations of 4 slices
2 pregnancy´s
3 hormonale activity
2 no hormonale activity
1 primary atrophy of the ovary
Cryopreservation of Ovarian Slices

43. Conclusion
Whole ovary: - technical possible
- major operative procedure
- endothelial cells of capillary system
- not ready for application in human
Slices: - easy surgical procedure
- minor risk
- repeatable
- relative and absolute reduced
follicle pool

44. Helga Bergmeister
Markus Lipovac
Gerda Hofstetter
Margareta Rudas
Christine Kurz
Rene Wenzl