Successful Uterus Cryopreservation in an Animal Model

Abstract

Cryopreservation of whole organs would not only be advantageous for experimental biology but also for transplantation surgery. The objective of this study was to evaluate the feasibility of cryopreserving swine uterus as an experimental model of cryopreservation affecting a whole visceral muscle organ. Organs were frozen slowly (0.2 °C/min) after arterial perfusion with 10 % dimethyl sulfoxide solution for 10 min and equilibration in this solution for 30 min. Viability of the organs was tested by biochemical parameters, LIVE/DEAD fluorescent staining and capability for contraction in a perfusion system. Ten fresh and ten cryopreserved uteri showed living cells in the LIVE/DEAD assay, and were viable for at least 7 h as shown by their ability to produce rhythmic contractions after oxytocin administration. This is the first report known to us on successful cryopreservation of a whole swine uterus to demonstrate that perfusion with a cryoprotectant and slow freezing provides a promising method for cryopreserving whole organs, and may encourage further studies with other cryoprotective agents and freezing protocols as well as other organs.

Female genital tract - ovary - perfusion - dimethyl sulfoxide - rhythmic contractions

References

• 1 Bottomley M J, Baicu S, Boggs J M, Marshall D P, Clancy M, Brockbank K GM, Bravery C A. Preservation of embryonic kidneys for transplantation.  Transplant Proc. 2005;  37 280-284
• 2 St Peter S D, Imber C J, Friend P J. Liver and kidney preservation by perfusion.  Lancet. 2002;  16 604-613
• 3 Jahania M S, Sanchez J A, Narayan P, Lasley R D, Mentzer R M Jr. Heart preservation for transplantation: principles and strategies.  Ann Thorac Surg. 1999;  68 1983-1987
• 4 Friedler S, Giudice L C, Lamb E J. Cryopreservation of embryos and ova.  Fertil Steril. 1988;  49 743-764
• 5 Pegg D E. The history and principles of cryopreservation.  Semin Reprod Med. 2002;  20 5-13
• 6 Rubinsky B. Principles of low temperature cell preservation.  Heart Fail Rev. 2003;  8 277-284
• 7 Dittrich R, Maltaris T, Mueller A, Dragonas C, Scalera F, Beckmann M W. The extracorporeal perfusion of swine uterus as an experimental model: the effect of oxytocic drugs.  Horm Metab Res. 2003;  35 517-522
• 8 Mazur P. Limits to life at low temperatures and at reduced water contents and water activities.  Orig Life. 1980;  10 137-159
• 9 Siebzehnrubl E, Kohl J, Dittrich R, Wildt L. Freezing of human ovarian tissue: not the oocytes but the granulosa is the problem.  Mol Cell Endocrinol. 2000;  169 109-111
• 10 Wang X, Chen H, Yin H, Kim S S, Lin T an, Gosden R G. Fertility after intact ovary transplantation.  Nature. 2002;  415 385
• 11 Guttman F M, Lizin J, Robitaille P, Blanchard H, Turgeon-Knaack C. Survival of canine kidneys after treatment with dimethyl-sulfoxide, freezing at -80 degrees C, and thawing by microwave illumination.  Cryobiology. 1977;  14 559-567
• 12 Picton H M, Gosden R G, Leibo S P. Cryopreservation of oocytes and ovarian tissue. In: Vayena E, Rowe P, Griffin PD (ed) Current practice and controversies in assisted reproduction. Geneva; World Health Organization 2002: 142-151
• 13 Maltaris T, Dimmler A, Mueller A, Binder H, Hoffmann I, Kohl J, Siebzehnruebl E, Beckmann M W, Dittrich R. The use of an open-freezing system with self-seeding for cryopreservation of mouse ovarian tissue.  Reprod Dom Anim. 2005;  40 250-254
• 14 Bedaiwy M A, Jeremias E, Gurunluoglu R, Hussein M R, Siemianow M, Biscotti C, Falcone T. Restoration of ovarian function after autotransplantation of intact frozen-thawed sheep ovaries with microvascular anastomosis.  Fertil Steril. 2003;  79 594-602
• 15 Bedaiwy M A, Falcone T. Ovarian tissue banking for cancer patients: reduction of post-transplantation ischaemic injury: intact ovary freezing and transplantation.  Hum Reprod. 2004;  19 1242-1244
• 16 Revel A, Elami A, Bor A, Yavin S, Natan Y, Arav A. Whole sheep ovary cryopreservation and transplantation.  Fertil Steril. 2004;  82 1714-1715
• 17 Imhof M, Hofstetter G, Bergmeister H, Rudas M, Kain R, Lipovac M, Huber J. Cryopreservation of a whole ovary as a strategy for restoring ovarian function.  J Assist Reprod Genet. 2004;  21 459-465
• 18 Martinez-Madrid B, Dolmans M M, VanLangendonckt A, Defrere S, Donnez J. Freeze-thawing intact human ovary with its vascular pedicle with a passive cooling device.  Fertil Steril. 2004;  82 1390-1394
• 19 Yin H, Wang X, Kim S S, Chen H, Tan S L, Gosden R G. Transplantation of intact rat gonads using vascular anastomosis: effects of cryopreservation, ischaemia and genotype.  Hum Reprod. 2003;  18 1165-1172
• 20 Maltaris T, Scalera F, Schlembach D, Hoffmann I, Mueller A, Binder H, Goecke T, Hothorn T, Schild R L, Beckmann M W, Dittrich R. Increased uterine arterial pressure and contractility of perfused swine uterus after treatment with serum from pre-eclamptic women and endothelin-1.  Clin Sci (Lond). 2005;  109 209-215
• 21 Racho El-Akouri R, Wranning C A, Molne J, Kurlberg G, Brannstrom M. Pregnancy in transplanted mouse uterus after long-term cold ischaemic preservation.  Hum Reprod. 2003;  18 2024-2030
• 22 Newton H, Aubard Y, Rutherford A, Sharma V, Gosden R. Low temperature storage and grafting of human ovarian tissue.  Hum Reprod. 1996;  11 1487-1491
• 23 Devireddy R V. Predicted permeability parameters of human ovarian tissue cells to various cryoprotectants and water.  Mol Reprod Dev. 2005;  70 333-343
• 24 Fageeh W, Raffa H, Jabbad H, Marzouki A. Transplantation of the human uterus.  Int J Gynaecol Obstet. 2002;  76 245-251
• 25 Altchek A. Uterus transplantation.  Mt Sinai J Med. 2003;  70 154-162