While adolescent and adult men have the option to cryopreserve sperm prior to chemotherapy or radiation in cancer treatment, this is not available to pre-pubertal boys. Researchers from the University of Pittsburgh School of Medicine and the Magee-Womens Research Institute write that several IVF centres worldwide are preserving testicular biopsies for these patients in anticipation that spermatogonial stem cells (SSCs) in the tissue can be used to achieve fertility in the future. In order to maximise the use of these tissues, the authors Valli-Pulaski et al, compared the cryopreservation of a cell suspension to an intact piece of tissue to discover the optimal cryopreservation method.
In the experiment the efficiency of each technique was analysed by immuno-cytochemistry for spermatogonia markers and human-to-nude mouse xenotransplantation. The average immuno-cytochemistry tested positive cells were highest in the fresh tissue; but from the cryopreserved groups, large tissue pieces and small tissue pieces had similar numbers of positive cells while cell suspension had the least positive cells and was significantly worse than cryopreserved intact tissue pieces. The authors conclude that based on their results, slow freezing of small piece of tissue is the most efficient technique to cryopreserve human testicular tissue.
For further information
Optimising Cryopreservation of Human Testicular Tissues
Reproductive Biomedicine Online
Volume 37, Supplement 2, Page e4
The development of an artificial liver has been of prominent interest, as liver disease is the fifth biggest cause of deaths in the UK. Acute liver failure (ALF) in children is a life-threatening condition. The prognosis of ALF is generally poor and medical intervention relies on a liver or hepatocyte transplantation if the native liver is unable to recover. The major limitation of the technique today is the availability of donor organs. There have been limited clinical advancements in the treatment of ALF utilising novel regenerative techniques, such as the use of embryonic stem cell techniques to induced pluripotent stem cell-derived hepatocytes. The use of human bio-compatible scaffolds to grow liver type/functional cells is increasing and the main challenges in translating advances in basic science of cell therapy into the clinic has been determining the best route of delivery, the rapid elimination of transplanted cells by the recipient, poor engraftment and proliferation of transplanted cells within the liver.
Researchers at Kings College London, Anil Chandrashekran (pictured right) and Anil Dhawan (pictured below), examined the use of methylcellulose as a scaffold to obtain liver-organoids. They cultured hepatocyte and MSCs in the presence of methylcellulose and growth factors and tested the ability of organoids to produce albumin and detoxify ammonium chloride, in optimally cultured conditions describing their research in BioInsights December 2018.
Mesenchymal stem/stromal cells (MSCs) can be easily cultured from adult bone marrow and full-term umbilical cord (blood or the Wharton’s jelly) and have been isolated from muscle connective tissue, adipose tissue and in some circumstances peripheral blood. Currently novel scaffolds are under scrutiny for the unmet need to use organoid-based product suitable for transplantation in ALF, this until the patient receives a liver transplant or recovers.
Hepatocytes fractions were collected, treated and cells were frozen in cryovials and cryobags using a Planer controlled rate freezer before storage at -180°C vapour phase nitrogen freezers. The MSCs were isolated from Wharton’s jelly of umbilical cords obtained following caesarean section deliveries. Once cell cultures were established, they were expanded, quality controlled and cryopreserved
The study showed that methylcellulose, an inert semi-solid media more suitable for clinical use than current products, could be utilised as a scaffold to establish liver-organoids that resemble liver structure and function. And they say the suitability of using methylcellulose towards clinical grade expansion of organoids is highly compelling. Further optimisation and scaling up of the process is underway.
Improvements to hepatocyte transplantation have been made by encapsulating hepatocytes with alginate beads which eliminates the need for immunosuppressants. More recently, bio-fabrication techniques have been developed using 3D plotting with methylcellulose and alginate. MSCs cultured in these 3D scaffolds retained viability and differentiation properties and so taking these techniques together it should be feasible to establish organoids in methylcellulose, as done here and then encapsulating the resulting organoids in alginate-methylcellulose 3D scaffolds. The organoid culture system established is thought highly applicable in the treatment of ALF but might also be well suited to drug screening and disease modelling.
For further information
Read article: Methylcellulose as a scaffold in the culture of liver-organoids for the potential of treating acute liver failure
Planer freezers: Medium sized control rate freezers
The 4th Upper Egypt Assisted Reproductive Conference (UEARS), will take place in Cairo from the 20th – 22nd February 2019 and is aimed at gynaecologists, andrologists, embryologists, lab directors, medical field researchers and those involved in Quality Control and Total Quality Management of the ivf laboratory.
This year, UEARS hopes to welcome over 2000 delegates and will offer a cutting-edge programme, with an international line up of speakers. UEARS’ goal is to foster innovation and collaboration and to fill the gap between science and practice.
Planer will be supporting our local distributor, Modern BioSystems. Come and see us on stand 7 to find out more about our CT37stax™ benchtop incubator, DATAssure™ laboratory wireless alarm and monitory system and our range of cryogenic freezers.
Research from a team at Nottingham University at the "Fertility 2019" conference presented some interesting results about uterine cryopreservation, which, considering recent reports on the success of uterine transplants, may be a viable option for uterine banking in the future. Author Dr Juan Hernandez-Medrano (pictured here) published a poster on the "Evaluation of tissue after uterine cryopreservation following a slow freezing protocol" where he was looking into possibilities for uterine freezing that could be an option for uterine factor infertility in patients. Currently, cold ischaemic storage is the preservation strategy between retrieval and transplantation. Cryopreservation offers the possibility of storage allowing a longer window between surgical procedures, provided tissue viability.
In the project, ovine uterine horns were dissected, perfused with cold heparinised saline followed by cryoprotectant and frozen using a Planer controlled-rate freezer with the method reported for ovarian cryopreservation by Campbell et al., 2014. The study showed that following cryopreservation, uterine tissue retains its functional characteristics (myometrial contractility and endometrial endocrine activity) but at a reduced rate though - potentially due to increased tissue damage as indicated by elevated lactate dehydrogenase release.
Dr Hernandez-Medrano mentions that this a pilot study and follow-up experiments are expected to improve on the above results which are encouraging and indicate that improvement of cryopreservation protocols taking account of infusion rate/time and the freezing rate might one day allow uterine tissue banking and transplantation in human patients.
This study was carried out in collaboration with Dr R Robinson from the Vet School at Nottingham University and Prof B Campbell. Konnary V, Robinson R, Campbell BK and Hernandez-Medrano JH. Evaluation of tissue after uterine cryopreservation following a slow freezing protocol. Fertility Conference 2019, ICC Birmingham, 3rd-5thJanuary 2019 https://tinyurl.com/ydggxdof
Fertility 2019, held in Birmingham on 3rd January, was the Joint Conference of the Association of Clinical Embryologists, British Fertility Society and the Society for Reproduction & Fertility which Planer were proud to attend showing their latest benchtop incubator and wireless alarm systems as well as the full controlled rate freezer range.
So to round off our 45th year in business, we have produced an illustrated profile which looks at our internal operations, history, product range and the role that our distributor family have played in our success.
At the same time we would like to thank colleagues and collaborators for their help since we started in 1973. Our company has evolved from pioneering one off machines for cryogenic researchers to becoming the gold standard supplier of equipment for cell preservation across a broad range of fields including IVF, cryobiology and stem cell research with a range of freezers, incubators and laboratory monitoring systems that help achieve many notable scientific breakthroughs.
The Ovarian Club is a unique meeting that brings together leading experts to share knowledge and experience in all facets of ovarian function. The meeting is a gathering of scientists, clinicians and embryologists who have a common interest in oocyte and early embryo development and the implantation process.
The 2018 meeting addressed the challenge of bridging science and clinical practice, focusing on ovarian function from oocyte recruitment to the stages of implantation and embryo development.
Prior to this meeting in Paris, a fertility preservation workshop was held under the direction of leading Fertility Preservation expert Professor Dror Meirow from Israel. The workshop was a hands-on clinical and laboratory program designed for medical doctors, embryologists and scientists who are interested in learning, discussing and practicing principals, clinical issues and different techniques of fertility preservation.
The workshop focused on the cryopreservation and transplantation of ovarian cortical tissue. Lectures focused on tissue harvesting, laboratory techniques, tissue transport, tissue evaluation and storing and documentation.
Ovarian Tissue Preservation is becoming well established as a viable technique for fertility preservation especially for pre-pubescent teenagers and patients undergoing cancer treatment. There have now been over 130 successful births worldwide. Cryopreservation, from controlled slow freezing, proving to provide the greatest chance of success with over 125 of the births resulting from ovarian tissue preserved using this technique.
The Planer Kryo 360 freezer has been at the forefront of the development of ovarian tissue cryopreservation and continues to be used by clinics throughout the world that are offering this option to their patients.
Since the launch of our CT37stax™ benchtop incubator in 2017, we have talked extensively to our distributors and customers, who have used the incubator, to get important feedback. Based on this information, our engineering team have been busy working on a number of improvements that are now ready to be released.
These new features will be fitted to all CT37stax™ models at no additional cost.
NEW – Soft closing incubator lids
We have added a soft close mechanism to the lid of each incubation chamber. This dampener will ensure the lid of the incubation chamber will always close smoothly even if accidentally knocked or dropped during manual closing.
NEW – Incubator Chamber Movement Controller
We have also added a mechanism that controls the speed of movement of the incubation chambers as they move in and out of the stack. The mechanism provides a smooth consistent speed ensuring that there are no sudden movement or forces applied to the embryo being cultured inside the chamber.
NEW – Improved Incubation Chamber Locking Mechanism
To reduce the risk of accidental disconnection of the incubation chamber from the stack, we have added an additional mechanical securing device to each chamber shelf to hold the chamber in place.
NEW - Reusable Humidification Water Trays
Each CT37stax™ incubator will be issued with a set of plastic humidification water trays which will last up to 4 weeks with regular refilling, however there is now the option to purchase stainless steel water trays that can be sterilised and reused.
We will of course continue to develop further improvements as more feedback is received from the market.
Find out more about the CT37stax™ benchtop incubator
Download the CT37stax™ benchtop incubator brochure
Download the CT37stax™ benchtop incubator specification sheet
Planer will be supporting our local distributor, IEPSA Medical Diagnostics, who will be exhibiting our latest CT37stax™ benchtop incubator and our DATAssure™ laboratory wireless alarm and monitory system at the Reproductive Medicine conference on the 23 – 25 November 2018 at the Ekurhuleni International Convention Centre in Johannesburg.
The conference’s main goal is to improve the reproductive health of all women from puberty to menopause. Discussions will be held on the latest developments in Assisted Reproductive Technology and its practical application in day-to-day clinical practice. This meeting will be a sharing of ideas on the current evidence-based investigation and management of fertility and the latest update on surgery to improve fertility.
Distinguished experts, from leading South African and international institutions, will be speaking. The conference presents an ideal opportunity to interact with colleagues and create lasting relationships.
Prior to the conference, IEPSA have organised a customer workshop covering a wide variety of topics including providing the optimum incubator environment, ensuring the safety of the samples in the ART clinic, Immunoassay Hormone Testing and Easy, Fast and Specific depletion of unwanted apoptotic spermatozoa.