News 2018

45 years - still pioneers

45 years logoPlaner plc celebrates 45 years
In 2018 Planer plc, formerly known as Planer Products, will be celebrating 45 years of designing, manufacturing, selling and supporting products for a variety of technical uses throughout the world. The company sprung from GV Planer Ltd., founded in the 1950s, primarily a design and research consultancy making one off products for research institutions. It was some of these earlier research products that were the foundations of the equipment that Planer plc went on to develop into commercially viable machines. These continued to evolve over the 45 years into the products we manufacture and sell today.

Slow freezing pioneers
The first birth from a frozen bovine embryo was reported in 1972 - using a very early Planer machine. Professors Rawson and Wilmut pioneering the animal husbandry field. With its successful development and the use of products for controlled rate freezing, the company received the Queen's Award for Technology and also awards from the British DTI for Innovation and Good Practice in Micro-electronics and achieved approval and for the demanding standards of medical device manufacture: ISO13485:2012 & Medical Devices Directive, Annex 11 93/42/EEC: LRQA. ISO 9001:2008.

The use of controlled rate freezers widens
IVF labs were the initial market but later hospitals and researchers found they needed programmable freezing for many types of cells – in areas such as cord blood banking, bone marrow transplants, botanicals, semen, oocytes, skin, ovarian tissue, heart valves and blood vessels. Planer Controlled Rate Freezers continue to be at the forefront of many of today’s scientific breakthroughs using cryopreserved cells.

Incubators
Forty two years ago in 1976 we conceived our very first incubator whilst collaborating with the then UK Department of Health and Social Services in the production of a new form of anaerobic incubator as an alternative to the prevailing jar system for the isolation of anaerobes from clinical material. The prototype machine was built around 1980 and was tested by a team led by Dr Berry from the Department of Microbiology at St Thomas's Hospital Medical School London, SE1 in 1981. The Planer range of CO2 incubators has since been developed further with the introduction of the very successful BT37 in 2009 and the new CT37stax multi chamber incubator in 2017.

Throughout 2018, we plan to run a series of articles and events to celebrate our 45 years of working with and supporting scientists and healthcare professionals who use our equipment to preserve, protect and nurture all types of cells in many important applications throughout the world.


Pictured here: Professor David Pegg with a Planer freezer in the 1970s.

 

 

 

 


 

Preservation of Cells: A Practical Manual by Allison Hubel

Professor Allison HubelA new book has just been published by Professor Allison Hubel of the University of Minnesota. It aims to provide readers interested in cryo bio banking with the tools needed to develop or debug a preservation protocol for cells. The core structure and content of the text grew from a professional short course that she has offered at the Biopreservation Core Resource over the last 10 years. The text describes, step by step, the individual elements of a protocol, including the relevant scientific principles for each phase. It can be used by anyone who is involved in cell preservation - including those not expert in the freezing of cells; it does provide the scientific basis for those that want to understand the basis for the protocol.

The book covers pre-freeze Processing and Characterization; Formulation and Introduction of Cryopreservation Solutions; Freezing Protocols; Storage and Shipping of Frozen Cells; Thawing and Post Thaw Processing; Post-thaw Assessment; and Algorithm driven Protocol Optimisation. Professor Hubel explains the reasons behind every step in the development of a preservation protocol and the scientific principles behind them and provides alternative modes of preservation for when conventional methods of cryopreservation are not appropriate for a given cell type or application.

Preservation of Cells A Practical Manual by Allison HubelMammalian cells have become modern workhorses capable of a variety of applications, such as in production of therapeutic proteins, viral vaccines, antibodies and therapeutic agents. Cells that are to be used therapeutically must be properly stored to meet the safety and quality control testing prior to release of the cells. There are a variety of methods that can be used to preserve cells depending upon downstream application.  Specific practices are intended to improve both the outcome and the reproducibility of preservation protocols. The preservation toolkit is used to develop fit-for-purpose preservation protocols. Each element of a protocol is based on scientific principles and those principles are explained and can be used to rationally design that particular step to achieve the desired downstream use of the cell. The influence of processing procedures on the health of cells prior to cryopreservation and strategies to mitigate stress on cells is explained along with the formulation and introduction of cryopreservation solutions.

There is a specific chapter on controlled rate freezing with all the steps of a controlled rate freezing protocol and guiding principles for design of that step are given. Also described is a process for transferring units from a controlled rate freezer to a storage unit and there are examples showing good and bad freezing curves. Prof Hubel also covers storage and shipping of frozen cells, processing, assessment, protocol optimisation and the cryopreservation of endothelial cells in suspension, peripheral blood mono nuclear cells from whole blood, human adipose stem cells, red blood cells, oocytes and more.

Preservation of Cells: A Practical Manual is an important book for researchers, laboratory technicians and students in cell biology, stem cell biology, tissue engineering, and regenerative medicine. It is also useful to cell bankers, regenerative medicine, biomarker discovery or precision medicine companies, and cell therapy labs, blood bankers, biobankers, and biotechnology companies.

For further information:
Prof Allison Hubel http://www.me.umn.edu/people/hubel.shtml
Preservation of Cells: A Practical Manual (John Wiley & Sons, Inc., Print ISBN: 9781118989845)
https://www.wiley.com/en-gb/Preservation+of+Cells:+A+Practical+Manual-p-9781118989845

 

 

 

Progress on a bio-artificial liver

A bio-artificial liver - a BAL - is used as an extra-corporeal organ designed to aid the function of the liver in patients with acute failure and University College London's team are working on one where liver cells are encapsulated in alginate. The challenge as a clinical product is related to immediate availability, because of the unpredictability of disease progression in acute liver failure.

The aim of a recent study, presented at the Society of Low Temperature Biology annual meeting by Eloy Erro, Joana da Silva (both pictured here) et al, was to achieve BAL biomass cryopreservation with different systems for post-thaw culture and recovery, aiming at an off-the-shelf product. HepG2 cells were encapsulated in 1% alginate solution, producing ~500µm beads which were encapsulated and cultured for 12-14 days in a fluidised bed bio reactor (FBB) to obtain sufficient biomass.

Cryopreservation of >1L of alginate cell beads was achieved in a cryo-bag using our controlled rate freezer Kryo 750 and DMSO was used as cryoprotectant with an ice nucleator.

The team concluded that successful cryopreservation of encapsulated liver cell spheroids on a large scale, at 16-48% of a total human liver size (the biomass of 30% liver cell numbers is the likely requirement for such a bio-artificial liver) represented the correct order of magnitude and provided sufficient post-thaw recovery. They also found the process could be easily scalable in freezing bags using the Kryo750 and the same fluidised-bed bioreactor for cell recovery. The use of micro-organoids within an alginate matrix proved an interesting choice for large-scale cryopreservation avoiding problems associated with large tissue freezing. However a more rapid recovery would be ideal, and the team is exploring improvements.

Further information:
Poster - The BAL Team : http://www.ucl.ac.uk/livergroup/theteam.html
Planer's controlled rate freezer: https://planer.com/products/cryo-freezers/medium-crf/kryo-750.html
To download PDF of poster, please click here.

bio artificial liver poster ucl

 

 

 

New controlled rate freezer at Cell Tech Pharmed

controlled rate freezers at CellTech Pharmed

Professor Nasser Agdami
Prof Nasser Aghdami, a user of Planer freezers when he was at Iran's Royan Institute, recently helped to set up a new company Cell Tech Pharmed to meet an increasing demand for stem cell therapy. The focus on several distinct areas - systemic diseases with an underlying inflammatory and immunologic etiology, cardiac and vascular diseases, orthopaedic diseases of the knee, ankle and pelvis where cell therapy products can be locally administered, improving outcomes of bone marrow transplantation and also skin augmentation where current methods are ineffective.

Embryonic stem cells or pluripotent stem cells can differentiate into any cell in the body. Because stem cells in the umbilical cord are less developed than adult stem cells, they don't have to be "matched" as stringently to a patient's human leukocyte antigen, or HLA, type. In the USA for example more than 80,000 unrelated bone marrow and umbilical cord blood transplants have been facilitated since 1987.

Embryonic stem cells or pluripotent stem cells can differentiate into any cell in the body. Because stem cells in the umbilical cord are less developed than adult stem cells, they don't have to be "matched" as stringently to a patient's human leukocyte antigen, or HLA, type. In the USA for example more than 80,000 unrelated bone marrow and umbilical cord blood transplants have been facilitated since 1987.

Planer Kryo 360 Controlled Rate Freezer at CellTech PharmedThe new company has the technical help and financial support of Dr Aghdami's old employer, the Royan Institute, along with the commercial backing of the Barakat Pharmed Company, a large pharmaceutical holding company.  In the pharma world, quality is of course paramount - the Barakat group complies with all common pharmaceutical standards such as the FDA, WHO, EMEA. Likewise in Cell Tech Pharmed who use one of our precision slow freezers, a Kryo 360, (pictured here) for their cell freezing.

 

Professor Brian Grout speaks at Stem Cell Users Group Meeting

We were pleased to be one of the commercial sponsors of this important meeting at Kings College Hospital, London, bringing together more than 60 stem cell practitioners to share knowledge and experiences and discuss current issues and difficulties. For this group, cryopreservation is a key enabling technology contributing to advances in both basic research and clinical practice. Consequently, a strong understanding of the principles underlying cryopreservation protocols is important to group members and we were happy to be able to support Professor Brian Grout*, a widely experienced cryobiologist and recent Chair of the Society for Low Temperature Biology, to contribute to the meeting.

Brian presented the audience with a cryobiologist’s view of the cryochain central to a stem cell user’s activities, moving from cell preparation to end use in the laboratory or on the clinical front-line. The intention was to present the pitfalls experienced by the audience in a cryobiological context and to point out where stronger biological understanding can be used to eliminate costly, and typically unintended, poor practice. Throughout, the concept was that all the cryopreservation team members need to be aware of the biological issues, especially where frozen material has to be recovered by separate groups e.g. clinicians, with the expectation of reproducible, high performance of the frozen product.

In particular, a good deal of attention was given to the benefits of controlled ice nucleation and the risks of damaging temperature rise to samples that are lifted up into the neck of storage vessels when searching for specific samples. Further, the potentially lethal effects of poorly controlled thawing protocols were examined together with some of the risks of microbial contamination of the cryogen in storage vessels and transit shippers.  The need for strong, supporting biological knowledge to underpin successful cryopreservation was underlined and also the continuing need for sustained research and developments in technology; this, to strengthen the level of control available to those using stem cell cryopreservation as a near-routine tool. Positive feedback from the meeting was strong and we are looking forward to continuing our links with this stem cell users group in the coming year.

*Emeritus Professor, Life Sciences, Copenhagen University

Evaluation of different protocols for the cryopreservation of mouse ovaries; T. Kolbe et al

Poster from the ISFP congress in Vienna November 2017
Institute of Laboratory Animal Science, University of Veterinary Medicine, Vienna

Evaluation of different protocols for the cryopreservation of mouse ovaries

Cryopreservation of gametes and embryos is a routine method for the long-term preservation of genetic material. The cryopreservation of ovarian tissue offers a valuable addition to these current approaches. The freezing of ovaries with subsequent thawing and surgical engraftment can restore fertility in young women undergoing cancer treatment with chemo- and/or radiotherapy and preserve important lines, strains or races of experimental, domestic and wild animals. During the last decade, several different protocols for the cryopreservation of mouse ovaries were published. In this study we compared four different protocols, which were mostly based on older protocols with some modifications (Liu et al., 2008; Migishima et al., 2003; Wang et al., 2009).

Summary
Ovarian cryopreservation is an easy but invaluable additional tool to archive mouse strains. In summary, all tested protocols used in this study resulted in good pregnancy rates, which did not significantly differ from the pregnancy rate of recipients transplanted with fresh ovaries. Ovarian tissue survived the aforementioned preservation techniques well and resumed follicle development and ovarian cycle after transplantation. Controlled freezing in straws with DMSO as cryoprotectant (protocol Gr. III) resulted in the best performance based on the fast resumption of reproduction and a generally high litter size.

 

Ovarian tissue cryopreservation slow freezing versus vitrification, Nikiforov et al

Dmitry Nikiforov At the world congress of the International Society for Fertility Preservation in Vienna in November important topics such as advances in ovarian cortex transplantation, assessment of ovarian reserve, in vitro follicle growth, medical protection from chemotherapy, reimplantation of ovarian tissue were on the agenda. Slow freezing of cryopreserved ovarian tissue (for fertility preservation in cancer patients) was a hot topic and there were a number of workshops and posters. One such - see below - was from Dmitry Nikiforov at the University of Teramo, Italy and also the Rigshospitalet in Copenhagen.

Dmitry is also an ESHRE/ReproUnion fellow in the Laboratory of Reproductive Biology. He has three years of clinical experience in human embryology and now is working on a doctoral project about ovarian tissue cryopreservation and in-vitro oocyte production from the tissue. At the Laboratory of Reproductive Biology in Copenhagen (under Prof Claus Yding Andersen) he is focusing on isolation of follicles from medulla tissue discarded during cryopreservation of ovarian cortex for further in-vitro culture. Another point of interest is to evaluate an effect of additional substances to the cryopreservation medium, which may improve survival rate of the tissue as well as improve surviving after implantation of the tissue back to the patient.

 

 

 

Ovarian tissue freezing

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:-  
https://planer.com/products/cryo-freezers/small-crf/kryo-360.html
C
ryopreservation of ovarian tissue for fertility preservation in young girls:-
https://planer.com/company/news/latest-news/149-latest-news/869-cryopreservation-of-ovarian-tissue-for-fertility-preservation-in-young-girls.html

For more information on the UCL courses at Louvain, email:
This email address is being protected from spambots. You need JavaScript enabled to view it.

 

 

News Stories - 2018

News Stories - 2017