The recent failure of two liquid nitrogen storage tanks at two separate facilities in the United States has highlighted the need to re-examine the approach to how reproductive laboratories handle the storage of tissue. To fully understand how to increase the security and safety of stored reproductive tissue, we also need to understand why tank failure occurs.
In their article, Kimball O. Pomeroy (pictured here) and Mark Marcon discuss how to manage the risks inherent with the storage of reproductive tissue, as well as where to store tissue, how to store them, how to reduce risks through quality control methods, and finally, options available for inventory management.
For further information
Download the full article ‘Reproductive Tissue Storage: Quality Control and Management/Inventory Software’.
Find out more about alarms and monitoring from Planer here.
Read more about Kimball Pomeroy and The World Egg Bank here.
In late 2018 Planer plc was awarded the contract for a DATAssure™ independent monitoring system in one of the largest IVF clinics in the Middle East, the Sidra Hospital, with its state of the art laboratory in Doha. The clinic expects to be responsible for around 15,000 IVF cycles per year.
The DATAssure™ system was installed in March 2019, when the Planer team had the task of installing over 200 sensors to many different types of equipment, including incubators, cryo tanks and heat plates. As the lab’s HVAC system was already online, the installation had to be completed without impacting the clean room environment and without causing any environmental changes to the VOC levels.
The installation was so large that it required two DATAssure™ base units and the Dialler unit. The first DATAssure™ base unit was solely for the numerous Planer BT37 benchtop incubators, while the second base unit was used to monitor all the other equipment within the lab. The installation was completed by two engineers over four days, despite much of the system configuration being done prior to shipment to the site.
On the final day, DATAssure™ training was delivered to the embryologists and lab staff who will be working at the facility. The system installed has created an intuitive monitoring system that is extremely easy to manage and expand if required. The Planer team left feeling very proud of the DATAssure™ system that they installed at the Sidra Hospital. Not only is this system one of the largest installations carried out by Planer, but that this state of the art facility now has a state of the art independent monitoring system with DATAssure™.
For further information
DATAssure™ laboratory wireless alarm and monitoring system
The 35th Annual Meeting of the European Society for Human Reproduction and Embryology (ESHRE) will be taking place in Vienna from 23rd – 26th June 2019. Over four days, experts in reproductive medicine and science will gather to attend a broad range of scientific sessions and to network with their peers. Last year, over 12,000 people from 130 countries attended.
This year’s meeting will cover a range of hot topics in the field, including presentations on fertility preservation following cancer and ovarian tissue cryopreservation and transplantation.
Come and see the Planer team on Stand E119
Planer will again be exhibiting at ESHRE this year. Do come and see us on Stand E119, to find out more about our CT37stax™ benchtop incubator, our DATAssure™ laboratory wireless alarm and monitoring system and our range of slow freezers.
A new paper published in Human Reproduction March 2019 helps to solve the question as to whether it is possible to eliminate metastasized cancer cells from ovarian cortex fragments prior to auto transplantation - without compromising the tissue or follicles. In ex vivo experiments pharmacological inhibition of YAP/TAZ enabled the team from Radboud University, The Netherlands, to eradicate from human ovarian tissue fragments induced small tumours. This research is important because auto transplantation of ovarian tissue fragments containing tumour cells could reintroduce malignancy to the recipient.
Human ovarian tissue was obtained after female-to-male sex reassignment surgery and human rhabdomyosarcoma, leukaemia, breast cancer and Ewing’s sarcoma cell lines were used to induce tumour foci. The tissue fragments were cultured to allow formation of metastasis-like structures followed by a 24 hours ex vivo treatment to eradicate cancer cells. Rhabdomyosarcoma and leukaemia cells were effectively purged from the cortex tissue but breast cancer and Ewing’s sarcoma did not respond. Ovarian tissue integrity seemed not affected by purging and no statistically significant difference was observed in the percentage of morphologically normal follicles, percentage of follicles with apoptotic cells, follicular viability or glucose uptake between the control treated ovarian cortex and Verteporfin treated ovarian cortex.
The results of the study, funded in part by Merck B.V., seems to show that ex-vivo tumour cell purging of ovarian cortex fragments for fertility preservation is feasible and helps the safety of ovarian cortex auto-transplantation, increasing the chances that this form of fertility restoration could become an option for patients with malignancies where ovarian cortex transplantation is currently considered unsafe. The team, led by Dr Callista Mulder, are unclear whether the results reflect the behaviour of malignant cells that have metastasized to the ovary during natural disease progression and suggest that the functionality of the ovarian tissue after ex vivo treatment requires further investigation in vivo.
For further information
Enhancing the safety of ovarian cortex autotransplantation: cancer cells are purged completely from human ovarian tissue fragments by pharmacological inhibition of YAP/TAZ oncoproteins
Callista L Mulder Lotte L Eijkenboom Catharina C M Beerendonk Didi D M Braat Ronald Peek
Human Reproduction, Volume 34, Issue 3, March 2019, Pages 506–518
Researchers at the University of Michigan working with mice report on a new approach to assist post cancer treatment girls; chemo and radio therapy for cancer are toxic to the follicles and normally leave patients sterile. Currently a young female patient’s route to pregnancy is to freeze ovarian tissue prior to treatment with the hope that follicle growth and maturation are a viable procedure.
In this case, the team, led by Lisa Green and Hong Zhou demonstrated they could improve the rate at which ovarian follicles developed mature eggs by surrounding the follicles with adipose derived adult stem cells in a 3D scaffold, mimicking the environment of the ovary. Historically, attempts to grow human follicles into eggs in two-dimensional petri dishes have failed. The new approach increased follicle survival from less than 5% to between 42% and 86%.
These 3D scaffolds allow a single follicle to grow in all directions within a polymer network - a hydrogel. By surrounding the follicle with the adult stem cells, the researchers created a delivery system for cytokines, growth stimulating substances that improved the chances for success.
If the process became a procedure, it would involve an ovary being removed and frozen prior to treatment and, after recovery and should pregnancy be sought, follicles would be isolated and grown in vitro using the adipose derived adult stem cells till the eggs matured. "Eventually translated to human application, the hope is ovarian follicles can be taken, grown in vitro, healthy eggs obtained and a normal IVF procedure followed,” said Associate Professor Ariella Shikanov, pictured here.
The 9th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2019) will be held on 2 – 5 May 2019 at the Hong Kong Convention and Exhibition Centre. This year, the annual congress is jointly organised by the Asia Pacific Initiative on Reproduction (ASPIRE) and the Hong Kong Society for Reproductive Medicine (HKSRM).
Over the four days, scientists, clinicians, nurses and embryologists will be coming together to share their expertise. In particular, the future of reproductive medicine will be discussed, dissected and presented during the pre-congress and main congress.
We are delighted to announce that Planer will be exhibiting at Aspire 2019. Do come and see us on Stand A44 to find out more about our CT37stax™ benchtop incubator, DATAssure™ laboratory wireless alarm and monitoring system and our range of slow freezers.
A combined team from Singapore's Sincere IVF and Gleneagles Hospital, report and describe successful ovarian freeze, transplant and ICSI fertilisation in the Journal of Reproductive Biotechnology and Fertility (Dr Jaffar Ali editor-in-chief). Ovarian tissue freezing is still a novel and complicated technique which can be used clinically for fertility preservation in female children, adolescents and adults with cancer.
The team, led by Professor SC NG, performed the auto-transplantation of cryopreserved ovarian tissue and reestablished a regular menstrual cycle, collecting one egg - and its successful fertilisation showed ovarian function had been restored. In the paper, published early 2019 the authors (from Embryonics International, ACS Clinic for Woman and Sincere IVF) tell that the patient, initially diagnosed with a serous borderline tumour for both ovaries, but with no definite stromal invasion, has, subsequent to the intervention and to date, been well with no evidence of relapse noted.
The 37-year old woman patient, whilst healthy, had twenty years absence of fertility due to her husband’s azoospermia; she was diagnosed with ovarian cancer and an oopherectomy was laparoscopically performed in 2016 at Gleneagles Hospital, Singapore. The subsequent investigation showed serous borderline tumour for both ovaries but with no stromal invasion.
Three slices of morphologically normal tissue from the right ovary were collected and immediately delivered to the laboratory where the medullary tissue was removed from the ovarian pieces, and a total of 10 pieces of ovarian cortex tissue approximately 5mm by 5mm and about 1mm in thickness were prepared for cryopreservation (see below). The slow freezing method (Andersen et al., 2008 https://www.ncbi.nlm.nih.gov/pubmed/18603535) was employed to freeze the prepared pieces of the ovarian tissue.
All ten pieces of ovarian tissue were equilibrated at 4 °C and were then placed into cryo-vials for computer controlled freezing with a Planer Kryo-360. The initial cooling rate was −2 °C/min to −9 °C with manual seeding performed and cooling was continued at different ramps to−140 °C, when the samples were plunged into liquid nitrogen at −196 °C, and transferred to a liquid nitrogen storage tank.
Neither chemotherapy nor radiotherapy was performed on the patient who ten months later underwent heterotopic transplantation of thawed ovarian tissue. Two cryo-vials containing four pieces of ovarian tissue were thawed for transplantation on 16 March 2017. The cryopreserved tissue was placed into a peritoneal pocket in the region of the broad ligament, below the fallopian tube. Menstruation commenced two months later after induction and follicle growth was monitored in the fifth menstruation cycle with one follicle observed in the peritoneal pocket.
The oocyte was successfully retrieved after the fifth cycle, six months after transplantation and fertilised via intracytoplasmic sperm injection (ICSI) using the husband’s frozen-thawed testicular sperm. One 7-cell cleavage stage embryo was then frozen but no embryo transfer has been performed to date.
Pictured here, Professor Soon-Chye Ng, one of the authors mentions "The results of our first ovarian tissue transplant confirmed previous reports and lent support to the objective that ovarian tissue freezing can preserve fertility for cancer patients. This success demonstrated that cryopreservation of ovarian tissue and subsequent transplantation is a viable option for preserving fertility."
Worldwide, it is estimated that several thousand women have had ovarian tissue cryopreserved, but the number of transplantations remains low although it is accepted in an increasing number of countries; to date, more than 100 children have been born from this procedure worldwide (Andersen et al., 2018 https://www.ncbi.nlm.nih.gov/pubmed/29718400).
The authors conclude that although cryopreserving of ovarian tissue for fertility restoration is still regarded as experimental, successful collection of oocytes and fertilisation from auto transplanted ovarian tissue, such as here, show that ovarian tissue freezing can preserve fertility for cancer patients. Goals for the future should include optimising the outcomes of transplantation, with improved tissue quality, reduced follicle loss, shorter transportation times, and longer periods of functionality in cryopreserved ovarian tissue. Such improvement in technique is important as survival rates for cancer patients in their early reproductive life continue to improve with advances in cancer treatment.
For further information
Read Paper http://www.ivf-hub.net/journal-archive-new/2019-2/
Planer Kryo 360: https://planer.com/products/cryo-freezers/small-crf/kryo-360.html
More from http://www.ivf-hub.net
The Bio-preservation Core Resource at the University of Minnesota is offering a new course on the emerging issues in preservation of cell therapies aligned with the fundamentals of cryo-preservation. The course includes cryo protocol development, the designing of a storage facility plus practical advice on cryo containers and equipment, reagents, regulatory issues along with clinical aspects of cryo-preservation and quality control.
The presentation will be given in person as a course on May 21st to 22nd, 2019 in Minneapolis with a simultaneous webcast of proceedings. The lecture topics will cover:-
The course material includes both fundamentals on preservation as well as emerging issues and approaches. The course is appropriate for managers of cell therapy laboratories, technicians who preserve cells as a part of their daily routine, scientists involved in the development of cell therapies, companies that produce products for the cell therapy space, repositories that store cells to be used therapeutically and related disciplines.
The class is offered in two different formats - via class attendance or via a simultaneous webcast of the class for those who cannot attend the in-class lecture. Lectures are recorded and attendees over the web have the opportunity to review the lectures at their convenience.