The fifth World Congress of the International Society for Fertility Preservation (ISFP) took place in November 2017 in Vienna, Austria. Attended by many delegates from around the world topics included Cryopreservation and Reimplantation of Ovarian Tissue, Oocyte and Embryo Freezing, the search for Cancer cells in the Ovaries. And of special interest were the workshops and particularly those given by Professor Christiani Amorim on the cryopreservation of Ovarian Tissue. These workshops included hands on lab and clinical aspects, tissue collection, lab preparation and tissue freezing, storing, thawing and transplantation techniques.

Cancer treatment can save a life but it can also bring infertility; chemo and radio therapy destroy cancer cells but they also destroy eggs and the gonadal toxicity results in early menopause and ovarian failure. UCL Louvain pioneered post-cancer female fertility restoration by freezing ovarian tissue prior to treatment and subsequently reimplanting it. Cryogenically slow freezing a patient’s own ovarian tissue prior to storage, treatment and then reimplanting it afterward has been successful with some 130 children born worldwide, including the world's first at UCL’s Saint-Luc University Hospital in 2004.

Professor Amorim was an Associate Professor at the Brasília University, in Brazil before moving to Belgium. In recent years, she has focused her attention on ovarian tissue transplantation for cancer patients. Her pioneering studies have served as the basis for establishing the field of ovarian tissue engineering and she organised the first study group on reproductive tissue engineering. In her Vienna workshop - which was oversubscribed - she went through and demonstrated the techniques of cryopreservation of ovarian tissue and the slow freezing method a process very much pioneered by UCL Louvain.

In the presentation, Professor Amorim summarised that there have been more than 130 live births using the slow freezing protocols in a controlled rate freezer. This contrasts with only 2 or 3 live births using the vitrification technique. UCL Louvain has itself had 14 live births since she and Professor Donnez first tried the technique.

Workshop on the cryopreservation of Ovarian Tissue
The workshop ran through the established Louvain procedure.


Ovarian tissue preparation
An ovarian biopsy is taken, tissue being retrieved from the operating theatre and then prepared in sterile conditions. It is treated with cryopreservatives before slow freezing and cryo storage. Analyses are carried out prior, to check suitability, testing with patient serum, microbiological analysis and via the anatomopathology lab


Preparing the cryovial
The treated tissue is placed in vials, marked and sealed and then frozen in a specific protocol in a controlled rate freezer - in this case a Planer Kryo 360.



Loading the vial in the controlled rate freezer
As can be seen from the protocol below, the vial holder is removed at a critical temperature and is seeded by touching with cold forceps. In this case it was undertaken at -8 Deg C. The computer controlled freezer cools the sample at a pre determined series of temperature rates and gives printable result for medical records and audit.


'Seeding' the vial with cold forceps



Ovarian tissue freezing cooling curve

Once the sample has been controlled frozen it is carefully removed and placed in frozen quarantine at liquid nitrogen temperatures awaiting test results. If all is correct, it is then transferred to a cryo storage tank for longer term storage at liquid nitrogen temperatures; when needed it is retrieved and warmed quickly in a water bath prior to use. It can be many years before the patient is ready to use the sample; in 2015 a Dubai patient had tissue frozen in 2001 thawed giving a successful transplant and a baby girl.

More information:-
Kryo 360 controlled rate freezers:-
ryopreservation of ovarian tissue for fertility preservation in young girls:-

For more information on the UCL courses at Louvain, email:
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