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Autologous Reimplantation of Whole Ovine Ovaries
Martin Imhof
Department of Biomedical SciencesMedical University of Vienna
Landesklinikum Weinviertel Korneuburg
Cryopreservation of Ovarian Tissue
Follicle loss after implantation
Limited follicle pool
Cryopreservation of Ovarian Tissue
Connection to vascular system
Immediate perfusion
Cryopreservation of Ovarian Tissue
Size of ovary - Cryopreservation
Major surgical procedure
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
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
Freezing Curve
2C/min to 0C
0C for 12 min.
1.6C/min to -9C
8 min at -9C
by 0.4C/min to -40C
to -150C at the faster rate of 10C/min.
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.
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.
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.
Experiment 1 - Whole Porcine Ovary
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.
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
Ovary
Vessels
Salpinx Uterus
Reimplantation of a Whole Ovary- Vessel Preparation -
Reimplantation of a Whole Ovary- Vessel Preparation -
Reimplantation of a Whole Ovary- Vessel Preparation -
Reimplantation of a Whole Ovary- Vessel Preparation -
arteryvein
Reimplantation of a Whole Ovary- Vessel Preparation -
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 -
Slow Freezing
2C/min to 0C
0C for 12 min.
1.6C/min to -9C
8 min at -9C
by 0.4C/min to -40C
to -150C at the faster rate of 10C/min.
Reimplantation of a Whole Ovary- Anastomosis -
operation microscope
performing vascular anastomosis
Transplantation of a Whole Ovary
- Anastomosis -
vessel anastomosis
Reimplantation of a Whole Ovary
- Perfusion -
Reimplantation of a Whole Ovary
- Perfusion -
Reimplantation of a Whole Ovary
Reimplantation of a Whole Ovary
Reimplantation of a Whole Ovary
Reimplantation of a Whole Ovary
Reimplantation of a Whole Ovary
Reimpl. + 545 Tage
Reimplantation of a Whole Ovary
small vessels with healthy endothelial cells
Reimplantation of a Whole Ovary
Reimplantation of Ovine Ovaries
- Results -
Reimplantation of Sheep Ovaries
- Results -
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
Cryopreservation of Ovarian Slices
Ovary
Cryopreservation of Ovarian Slices
Radiation of Remaining Ovary
Radiation of Remaining Ovary
Reimplantation
Kontralaterales
Ovar
8 reimplantations of 4 slices 2 pregnancy´s 3 hormonale activity2 no hormonale activity1 primary atrophy of the ovary
Cryopreservation of Ovarian Slices
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
Helga Bergmeister
Markus Lipovac
Gerda Hofstetter
Margareta Rudas
Christine Kurz
Rene Wenzl
