Review our most recent research publications and current grant information.

Asymptote is actively involved in cutting-edge cryopreservation research, both in order to improve our products as well as helping our partners get the most from our services. Below is a selection of publications related to cryopreservation and use of our equipment, as well as a summary of current research grants we hold.

General Research Papers

Bioprocessing for Cell-Based Therapies

The recent Bioprocessing for Cell-Based Therapies publication explores the very latest techniques and guidelines in bioprocess production to meet safety, regulatory and ethical requirements, for the production of therapeutic cells, including stem cells.

Dr John Morris contributed to the chapter concerning the Fundamental points to consider in the cryopreservation and shipment of cells for human application.

Stacey GN, Healy L, Man J, Hunt CJ, Morris J. Fundamental points to consider in the cryopreservation and shipment of cells for human application. In Bioprocessing for cell based therapies. Ed CJ Connon, Wiley Blackwell 2017, pp 167-185 ISBN: 9781118743416.

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  • Kilbride P, Lamb S, Milne S, Gibbons S, Erro E, Bundy J, Selden C, Fuller B, Morris J. Spatial considerations during cryopreservation of a large volume sample. http://dx.doi.org/10.1016/j.cryobiol.2016.05.013

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  • Fonseca F, Meneghel J, Cenard S, Passot S, Morris GJ. Determination of intracellular vitrification temperatures for unicellular micro organisms under conditions relevant for cryopreservation. PLoS ONE 11(4): e0152939. doi:10.1371/journal.pone.0152939;2016

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  • Kilbride P, Gonzalez-Molina J, Maurmann N, Mendonça da Silva J, Gibbons S, Selden C, Fuller B, Morris J. Impact of Storage at -80°C on Encapsulated Liver Spheroids after Liquid Nitrogen Storage. BioResearch Open Access. June 2016, 5(1): 146-154. doi:10.1089/biores.2016.0017.

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  • Clarke A, Morris GJ, Fonseca F, Murray BJ, Acton E, Price HC. A Low Temperature Limit for Life on Earth. PLoS ONE 8(6): e66207. doi:10.1371/journal.pone.0066207

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  • Massie I, Selden C, Hodgson H, Fuller B, Gibbons S, Morris GJ. GMP Cryopreservation of Large Volumes of Cells for Regenerative Medicine: Active Control of the Freezing Process. Tissue Engineering Part C Methods. DOI:10.1089/ten.TEC.2013.0571;2014

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  • Morris GJ, Acton E. Controlled ice nucleation in cryopreservation – A review. Cryobiology 66:85–92;2013

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  • Morris GJ, Acton E, Murray BJ, Fonseca F. Freezing injury: the special case of the sperm cell. Cryobiology 64:71-80;2012

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  • Murray BJ, Broadley SL, Morris GJ. Supercooling of water droplets in jet aviation fuel. Fuel 90:433-435;2011

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  • Gosden RG, Yin H, Bodine RJ, Morris GJ. Character, distribution and biological implications of ice crystallization in cryopreserved rabbit ovarian tissue revealed by cryo-scanning electron microscopy. Human Reproduction 25:470-478;2010

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Research papers featuring automated thawing systems

  • Initial performance data on the CellSeal® Automated Thawing System (paper in preparation).

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Research papers featuring VIA Freeze equipment

  • A scale down process for the development of large volume cryopreservation. Peter Kilbride G. John Morris, Stuart Milne, Barry Fuller, Jeremy Skepper, Clare Selden. Cryobiology 69, 367-375, 2014

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  • Massie I, Selden C, Hodgson H, Fuller B, Gibbons S, Morris GJ. GMP Cryopreservation of Large Volumes of Cells for Regenerative Medicine: Active Control of the Freezing Process. Tissue Engineering Part C Methods. 01/14; DOI:10.1089/ten.TEC.2013.0571

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Research papers featuring the EF600

Laboratory based cryopreservation:

  • TF Whale, BJ Murray, D O’Sullivan, NS Umo, KJ Baustian, JD Atkinson, and GJ Morris. A technique for quantifying heterogeneous ice nucleation in microliter supercooled water droplets.doi:10.5194/amtd-7-9509-2014

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  • Ryan MJ, Kasulyte-Creasey D, Kermode A, San SP; Buddie, AG. Controlled Rate Cooling of Fungi Using a Stirling Cycle Freezer. Cryoletters 35:63-69;2014.

  • Rastoll MJ, Ouahid Y, Martin-Gordillo, Ramos V, Vasconcelos V, del Campo FF. The development of a cryopreservation method suitable for a large cyanobacteria collection. Journal of Applied Phycology. Online Feb 2013

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  • T. Pacchiarini, C. Sarasquete, E. Olague, M.P. Herráez, E. Cabrita Transplantation of cryopreserved testicular germ cells in flatfish species: a useful technique for reproductive biotechnology. Cryobiology 65:343-344;2012

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  • Massie I, Selden C, Morris J, Hodgson H, and Fuller B. Cryopreservation of encapsulated liver spheroids using a cryogen-free cooler: high functional recovery using a multi-step cooling profile. Cryoletters 32:158-165;2011

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  • Morris GJ, Acton E, Collins D, Bos-Mikich A, de Sousa P. Optimisation of current Good Manufacturing Practice (cGMP) compliant controlled rate freezing for human embryonic stem cells. Cryobiology 61:406;2010

  • Morris, G.J., Acton, E., Faszer, K., Franklin, A., Yin, H., Bodine, R., Pareja, J., Zaninovic, N., and Gosden, R. Cryopreservation of murine embryos, human spermatozoa and embryonic stem cells using a liquid nitrogen-free, controlled rate freezer. Reproductive BioMedicine Online 13:421;2006

  • Morris, J. Power compensation analysis of cryopreservation samples during cooling in an electrically powered controlled rate freezer. Cryobiology 57;321:2008

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  • Smith D, Ryan M. Implementing Best Practices and Validation of Cryopreservation Techniques for Microorganisms. Scientific World Journal. Published online 2012 May 2.

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EF600 as a portable device:

  • Blanco J, Eason D, Vercoe D, Adams SL, Gale S, Moorhouse R. Cryoconservation in the field: the challenges of saving the Kakapo. Crybiology 61:391;2010

  • Faszer K, Draper D, Green JE, Morris GJ, Grout BWW. Cryopreservation of horse semen under laboratory and field conditions using a Stirling cycle freezer. Cryoletters 27:179-184;2006

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  • MartMaria S, DinisT, Saraquete C, Cabrita E. Biofreezer Vs. Nitrogen: New methods for European seabass (Dicentrarchus labrax) sperm biobanking. AQUA 2012 - Meeting Abstract

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EF600 as an analytical tool:

  • Atkinson JD, Murray BJ, Woodhouse MT, Whale TF, Baustian KJ, Carslaw KS, Dobbie S, O’Sullivan D, Malkin TL. The importance of feldspar for ice nucleation by mineral dust in mixed-phase clouds doi:10.1038/nature12278 (2013)

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  • O'Sullivan D, Murray BJ, Malkin T L, Whale T, Umo NS, Atkinson J D, Price HC, Baustian KJ, Browse J, Webb ME. Ice nucleation by soil dusts: Relative importance of mineral dust and biogenic components, Atmos. Chem. Phys. Discuss., 13, 20275-20317, 10.5194/acpd-13-20275-2013, 2013.

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  • Herbert RJ, Murray BJ, Whale TF, Dobbie SJ, Atkinson JD. Representing time-dependent freezing behaviour in immersion mode ice nucleation. Atmos. Chem. Phys. Discuss., 14, 1399-1442, 10.5194/acpd-14-1399-2014, 2014.

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Current Grants

Development of a regulatory compliant cassette for large volume cell culture, cryopreservation, thawing and perfusion.

This application relates to the development of the consumables and associated equipment to allow the widespread clinical delivery of a bioartificial liver (BAL). Since the liver is one of the few organs that can repair and regenerate, therapies enabling regenerative medicine, that is creating living functional tissues to repair or replace organ function lost due to damage, are expected to play a role in several areas of liver disease. A bioartificial liver machine can temporarily replace the functions of the liver, allowing the damaged liver to regenerate whilst protecting the patient’s other organs from the life-threatening damage that ensues during liver failure. If the toxicity can be mitigated, within 24 to 48 hours, the majority of liver cells will enter DNA synthesis, closely followed by mitosis enabling the liver to regenerate, restoring full function within a few days.

Innovate UK, Supporting regenerative medicines and cell therapies.

3 Years starting 01-05-2014

Equipment for shipping cryopreserved T cell therapies.

Cancer treatments are being transformed by T-Cell therapies and present a huge global opportunity. However, logistical problems transporting the cells to and from the patient are restricting the industry’s growth. There is an urgent need for a portable shipping device that combines; controlled sample freezing, temperature controlled shipping, data logging, short term frozen storage and thawing.

This project will develop a basic prototype, free of liquid nitrogen, optimised for the shipping of T-cell therapies. This will be an electrically powered system based on a Stirling cryocooler, with a target isothermal hold temperature of -120°C. The system will be able to operate on mains power, a 12V vehicle supply or via an uninterruptable power supply (UPS). The storage chamber will be vacuum insulated and the device will maintain -100°C for 24 hrs when disconnected from all power.

To allow transport of the source T-cells, the equipment will also be able to carry out the controlled rate freezing (CRF) of samples. The equipment will also act as a storage device at the clinical site, maintaining the sample temperature below -100°C until required for treatment. In discussions with a range of end users (academic and commercial) we have confirmed that there is currently no suitable cryogenic service for autologous treatments which can offer CRF of the source cells, temperature controlled transport, data logging, short term storage and thawing.

Innovate UK, SMART Proof of concept.

Nine months starting 15-09-2015

Improved methods for freeze drying of entomopathogenic fungi

The project brings together the technological expertise to develop a cost effective mass production and delivery a more effective biological solution to control pests with expertise in fungal cell processes and whole organism survival to ensure product long term shelf life whist retaining organism function. It combines improved product formulation with effectiveness to reduce crop losses and chemical pollution causing soil quality deterioration. The project will: Apply advanced technology to biological product development with the potential for transfer to other biological applications; Take improved laboratory knowledge to improve the cost effectiveness and efficacy to a product in the field; Develop formulations increase shelf life and confidence in the use of biological solutions to replace chemical pesticides; Produce a product appropriate for storage and use in developing economy countries; Reduce crop losses by utilisation of organisms that previously could not applied in the field.

This project will develop a basic prototype, free of liquid nitrogen, optimised for the shipping of T-cell therapies. This will be an electrically powered system based on a Stirling cryocooler, with a target isothermal hold temperature of -120°C. The system will be able to operate on mains power, a 12V vehicle supply or via an uninterruptable power supply (UPS). The storage chamber will be vacuum insulated and the device will maintain -100°C for 24 hrs when disconnected from all power.

Innovate UK, Materials for extreme environments

2 years starting 01-11-2015

Shear-thickening fluids for cryopreservation

In regenerative and transplantation medicine medicine, a bottleneck limiting progress is that tissue engineered constructs cannot be manufactured on demand. Cryopreservation aims to overcome this problem, however whilst success has been achieved with cell suspensions, successful scale up of construct size has remained elusive. No methods exist that can protect complex biomasses from the severe stress they encounter during cooling and warming from liquid nitrogen (-196°C). We propose a new method, where non-Newtonian, shear-thickening fluids can be used to improve operational performance of cryopreservation. Shear thickening fluids are materials whose viscosity increases with shear stress, for example vibration. With the correct level of vibration, the material can change from a liquid to a solid instantly. We propose this as an effective material for extremely low temperature biological preservation. At the storage temperature, shear stress would be stopped as the material would remain solid (vitrified) due to the low temperatures. The shear-thickening materials make the process completely reversible.

Innovate UK, Materials for extreme environments

1 years starting 01-06-2016