Ovarian tissue cryopreservation and transplantation techniques are rapidly becoming an accepted path for fertility preservation of prepubescence girls and women that require cancer treatment that would otherwise leave them unable to have families of their own.
To date, more than 130 children worldwide have been born using this new technique with by far the majority of these successful outcomes resulting from tissue frozen using a slow freezing process in a controlled rate freezer.
In our recently published “Ovarian Tissue Cryopreservation Scientific Round-Up” two different procedures are described - the first outlines the work from a combined team from Singapore’s Sincere IVF and Gleneagles Hospital whilst the second gives an overview of the approach used by Belgium’s Université Catholique de Louvain.
Ovarian tissue freezing has been used clinically for fertility preservation in children, adolescents and adults with cancer since 2004, following the world’s first live birth using the cryopreservation and transplantation of ovarian tissue.
Whilst the procedure has been accepted in an increasing number of countries, it is still a relatively new procedure, within the area of assisted reproduction technologies.
Why use ovarian tissue cryopreservation and transplantation?
Ovarian tissue cryopreservation represents a new technique to preserve fertility in women where these normal fertility preservation techniques are not an appropriate option.
Currently, embryo and oocyte cryopreservation are the only fertility preservation techniques considered by the American Society for Reproductive Medicine not to be classified as experimental. Unfortunately, there are several reasons why one of these approaches may not be appropriate for some patients:
For these patients, the possibility of cryopreservation of ovarian tissue (cortex) has become an urgent and highly-demanded technology.
Ovarian tissue cryopreservation and transplantation does not require ovarian stimulation and offers a promising option for women at high risk of premature ovarian failure and sterility. Furthermore, transplanting ovarian tissue not only restores fertility but also restores endocrine function.
Slow freezing versus vitrification
Ovarian tissue freezing and transplantation, using a controlled rate freezer, has a number of advantages over vitrification. The automated process of a programmable freezer allows repeatability, with an auditable trail, hence reducing the risk of human error. Slow freezing also allows larger grafts of ovarian tissue, which could be important in improving success rates.
Download our Ovarian Tissue Cryopreservation Scientific Round-Up
To find out more about the combined team from Singapore’s Sincere IVF and Gleneagles Hospital and the Université Catholique de Louvain’s approaches, download our Ovarian Tissue Cryopreservation and Transplantation Scientific Round-Up.
For further information
Ovarian tissue freezing
Successful oocyte retrieval and fertilisation after transplantation of cryopreserved ovarian tissue
New ovarian tissue freezing programme
Cancer cells purged from human ovarian tissue fragments by pharmacological inhibition of YAP/TAZ oncoproteins
First birth in Italy from cryopreserved ovarian tissue transplanted to a cancer patient
Increased follicle survival in frozen–thawed human ovarian tissue
First British woman gives birth after an ovarian tissue transplant
Unique birth from ovarian tissue frozen in 2001
We are delighted to announce that Planer is now the UK distributor for the Gynemed range of media products. If you are going to Fertility 2020, do drop by Stand One, to find out more.
The Fertility 2020 Conference, organised by the Association of Clinical Embryologists, British Fertility Society and the Society for Reproduction and Fertility, will take place in Edinburgh’s International Convention Centre from the 9th to the 11th January. The theme this year is “Reproduction in changing world” and will feature speakers from around the world, talking on fertility and reproductive biology.
Planer Ltd is now part of the Hamilton Thorne Group and we are now the UK distributor for the Hamilton Thorne Clinical and Research Laser Range, along with the Gynemed range of consumable and media products.
While at the Conference, come and talk to the Planer team on Stand One to find out more about Gynemed Media, designed for:-
At Planer, we are frequently asked the question “How do we optimise our freezing profile for our controlled rate freezer to get the best results?”. We thought that a summary of a recent presentation by Allison Hubel, might provide some useful guidance. Allison, an expert in Cryobiology from the University of Minnesota, USA, gave an informative talk to the Stem Cell users Group and the Society of Low Temperature Biology meeting in Seville where she explained the key factors to consider when trying to optimise a freezing profile. Here is an outline of her presentation on "Optimising Freezing Profiles"
Cryopreservation of a cell therapy
Cells therapies are becoming a standard of care for the treatment of disease and injury. Unlike other standard therapies (i.e. drugs and medical devices), cell therapies have a complex supply chain that requires viable, functional cells all along it. It is noteworthy that what happens along the supply and processing chain determines the quality of the product at the end. Cryopreservation of a cell therapy is a common method of stabilisation along this complex supply chain.
Cryopreservation typically requires 6 steps:
(1) Pre-freeze processing
(2) Formulation and introduction of a cryopreservation solution
(6) Post thaw characterisation.
Optimizing the freezing process
It has been known for almost 50 years that the rate of cooling has a strong influence on the post thaw recovery of cells. Controlled rate freezing is often used to control the temperature as a function of time for cells being cooled; therefore improving both the recovery and consistency of outcome.
A controlled cooling rate protocol has 5 steps:
(1) Initial equilibration
(3) Seeding of the sample
(4) Secondary cooling
(5) Cooling to the final temperature.
The initial equilibration stage of the freezing protocol enables samples placed in the controlled rate freezer to equilibrate with the freezer. Optimising this step, in particular, helps to improve reproducibility of the freezing process. The cooling rate for the sample is the cooling rate used for both primary and secondary cooling of the sample. One degree C/min is a cooling rate commonly used for several cell types.
Controlling the temperature at which ice forms in the extracellular solution
The temperature at which ice forms in the extracellular solution also plays an important role in the post thaw survival of cells. There are several methods of controlling the temperature at which ice forms in the extracellular solution: manual and automatic seeding. Manual seeding requires the use of liquid nitrogen or a chilled instrument to induce nucleation and this approach is commonly used with cell types that require precise control of the nucleation temperature. Automatic seeding uses a dip in the sample temperature to induce nucleation. Ultimately, the sample is cooled to a final temperature at which time it is transferred to a low temperature storage unit.
Debugging your freezing protocol
It is important to note that debugging your freezing protocol is straightforward; you can at any time stop the process, thaw the sample and determine viability. This approach allows us to determine if a particular segment of the controlled rate freezing protocol results in cell losses and a basis from which to change and optimise the protocol. Developing strategies for optimising and debugging freezing protocols will be critical in the development and implementation of high efficiency cryopreservation protocols.
Allison's presentation at the 2019 SLTB meeting was extremely well received and generated some thought-provoking discussions during the meeting. For more information, her book "Preservation of cells. A practical manual" provides an extremely useful guide to explore the subject in greater detail.
For further information
More about Allison Hubel
"Preservation of cells. A practical manual" by Allison Hubel
BioCoR, University of Minnesota, and the science, technology and practice of bio preservation
Society of Low Temperature Biology
Liver disease kills more than 12,000 people in the UK each year. As deterioration from liver disease is extremely rapid, any cell therapies to treat the disease must be readily available. A bioartificial liver machine (BAL) can provide the liver function and allows patient livers to regenerate.
The team at University College London have developed a BAL based on alginate encapsulated liver spheroids (AELS) and cryopreservation in >1 litre cryobags. The cryopreservation method enables long term storage and fast delivery of BAL to patients.
At the recent Society of Low Temperature Biology meeting in Seville, the UCL team presented a poster “Does DMSO toxicity in cryopreservation of liver cell derived organoid culture?”. Their approach is intended to help optimise a process for large scale biomass cryopreservation, suitable for a clinical bioartificial liver machine.
The team concluded that AELS can be exposed to 12% dimethyl sulfoxide (DMSO) for up to 10 minutes in temperatures of up to 37 °C without compromise to their viability of viable cell number. However, high concentration DMSO (40%) is toxic for the cells and significantly affects cell viability and cell number immediately after treatment after only a short exposure.
For further information
To download the poster, please click here
Progress on a bio-artificial liver
Cryopreservation for the UCL bioartificial liver
The Liver Group Charity
Society for Low Temperature Biology (SLTB)
Two years ago this November, baby Emma was born to Benjamin and Tina Gibson. The embryo that became Emma is, as far as is known, the longest frozen one resulting in a successful birth; it had been cryopreserved for more than 24 years prior to being thawed and transplanted.
The embryo was frozen in October 1992 and was thawed by the National Embryo Donation Center (NEDC) Lab director Carol Sommerfelt in March 2017. After frozen embryo transfer (FET) was performed by NEDC Medical director Dr Jeffrey Keenan, Tina Gibson 26 years old at the time, became pregnant with the donated embryo; it had been conceived just one and half years after her own birth!
"It is deeply moving and highly rewarding to see that embryos frozen 24.5 years ago using the old, early cryopreservation techniques of slow freezing on day one of development at the pronuclear stage can result in 100 percent survival of the embryos with a 100 percent continued proper development to the day-3 embryo stage," said Carol Sommerfelt.
At that time the Planer 1.7 freezer was universally used for embryo freezing, although it’s use here cannot be definitely confirmed after twenty-five years. In the 1990s most IVF centres cryopreserved embryos via the slow freezing Menezo method and used a Planer machine.
The NEDC runs a leading embryo adoption program, with nearly 900 pregnancies to its name. It helps utilise frozen embryos that would not otherwise be used by their genetic parents to assist sub fertile couples via donated embryo ‘adoption’.
National Embryo Donation Center www.embryodonation.org
Blastocyst Cryopreservation, 1995 paper: Kaufmann, Menezo et al https://www.fertstert.org/article/S0015-0282(16)57972-2/pdf
19-year frozen embryo birth https://planer.com/company/news/news-2013/273-2013-2/598-birth-from-an-embryo-frozen-19-years.html
The 6th World Congress of the International Society for Fertility Preservation (ISFP) will be taking place on 14th – 16th November in New York, USA. Over the past twenty years the demand for fertility preservation has grown dramatically – with the goal of improving the quality of life of young cancer survivors and helping those wishing to postpone reproduction due to medical or non-medical conditions. At ISFP this year, there will also be a pre-congress course on the cryopreservation of ovarian tissue, which will include a number of practical sessions.
Planer is delighted to be supporting and exhibiting at this year’s congress. Come and meet the Planer team to find out more about our range of products:
To find out more
6th World Congress of the International Society for Fertility Preservation (ISFP)
Planer Kryo 360 programmable freezers
Download our Kryo 360 specification sheet
ShipsLog3™ temperature data logger
DATAssure™ wireless monitoring system
The Middle East Fertility Society’s (MEFS) meeting will be taking place this year from 31st October to 2nd November in Cairo at the InterContinental Cairo Citystars hotel. Over the three days, experts from around the world will be speaking on broad range of themes, highlighting the latest in reproductive medicine.
If you are going to MEFS, do come and talk to the Planer team – we will be on the Modern Biosystems’ booth. Find out more about our range of products including our DATAssure™ laboratory wireless alarm and monitoring system, which can help monitor and prevent the accidental loss of samples, our compact CT37stax™ benchtop incubator and our selection of controlled rate freezers.
The Ovarian Club XIV Congress will be taking place in Paris from the 7th-9th November at the Westin Paris – Vendôme. The Ovarian Club brings together scientists, clinicians and embryologists who have a common interest in all facets of ovarian function, including oocyte and early embryo development and the implantation process. At this year’s meeting there Is also a pre-congress fertility preservation workshop, chaired by Professor Dror Meirow, with a particular focus on ovarian tissue cryopreservation and transplantation.
Planer is delighted to support this year’s Ovarian Club meeting. Come and talk to our team to find out more about:-