Autologous Reimplantation of Whole Ovine Ovaries

Martin Imhof

Department of Biomedical SciencesMedical University of Vienna

Landesklinikum Weinviertel Korneuburg

martin.imhof@meduniwien.ac.at

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