Cell culture technology
Recent advances, future prospects
9th March 2012
5th Annual event Friday, 09 March 2012
In the last 4 decades mammalian cell culture has matured from being merely a research tool into being one of the foundations of the biopharmaceutical industry, and its use is continuing to expand rapidly. In vitro models are replacing animals in many tests and assays; its enormous potential in the fields of stem cell and regenerative medicine has hardly started to be realized; and its utility in research grows ever faster”.
Meeting Chair: Dr John Davis, Chairman of the UK Branch of the European Society for Animal Cell Technology.
This conference will examine some of the latest applications of cell culture technology, some that are still “over the horizon”, and some of the problems that must be solved before it can reach its full potential.
This event has CPD accreditation and will have a troubleshooting panel session.
On registration you will be able to submit your questions to the panel that will be asked by the chair on the day of the event
9:00 – 9:45 Registration
9:45 – 10:00 Introduction by the Chair: Dr John Davis, Chairman of the UK Branch of the European Society for Animal Cell Technology
10:00 – 10:30 Polymeric matrix construction
Professor Alexander M. Seifalian, University College London, UK
10:30 – 11:00 A discussion of the systems used to measure autophagy in vitro
Katy Petherick, University of Bristol, UK
Autophagy is a rapidly growing field of research. Methods to analyse autophagy are constantly advancing. The complexity of the process makes it important to have accurate readouts of autophagic activity. This talk will discuss a range of methods for measuring autophagy and important points to consider when using these techniques.
11:00 – 11:30 Clinical grade human embryonic stem cells
Dr Dusko Ilic, Kings College London School of Medicine
hES cells are undifferentiated cells derived from an early embryo that can grow in vitro indefinitely, while retaining their capability to differentiate into specialized somatic cell types. Their use in therapy and regenerative medicine as well as in toxicity screening and drug development is widely anticipated. However, even if we pay no attention to ethical, religious and political issues that relate to hES cells, there are still a number of obstacles to be resolved before these cells can be broadly used for cell-based therapy. The talk provides an overview of the current status and the future perspective of the field from the point of view of the standard level of patient safety and efficacy for the healthcare industry.
11:30 – 12:00 Speakers’ photo then mid-morning break and trade show
12:00 – 12:30 Single use bioreactor for clinical-grade production of stem cells
Dr. Thorsten Adams, Sartorius Stedim Biotech GmbH, Germany
As the field of regenerative medicine further develops, there is an increasing need for larger scale generation of stem cell derived therapeutics for clinical applications. Stem cells are frequently cultured in simple devices such as petri dishes and static bags. However, the main drawback of these technologies is the lack of scalability and process control. Single use bioreactors are well suited for this application. They offer a controlled environment for hypoxic cultivations and the possibility for process automation. This talk will address the technology, first results and the challenges that need to be overcome in order to optimize the single use bioreactor technology for the regenerative medicine market.
12:30 – 13:00 Automated Approaches to the Optimisation of Stem Cell Expansion and Differentiation Professor Gary Lye, University College London, United Kingdom
Stem cell culture is a largely manual process, with major challenges to address in the methods used regarding scalability and variability. Process automation can be of great benefit to reduce operator-dependent variation, therefore improving cell yield and quality. This would be beneficial for production of defined cells for high throughput screening or definition of a robust cGMP process suitable for scaled-out production of cells for clinical application. This work describes the use of a custom assembled Tecan platform for the hands-free expansion and directed differentiation of a range of stem cells.
Key bioprocess variables were initially optimised to develop a Standard Operating Procedure (SOP) for the expansion and differentiation steps. Comparisons between the manual and automated process over eight sequential passages were then performed. Automated culture was shown to improve the consistency of cell yield up to 3-fold. Using the platform’s ability to control oxygen tension, stem cells were further differentiated into neural precursors at 2% oxygen and results compared to manual differentiation at 2% and 20% oxygen. Use of the enclosed automated platform avoided changes in oxygen tension during media changes, as occurs in manual culture. Up to a 16-fold increase in gene expression in the differentiated cell lines were obtained.
13:00– 14:00 Lunch and trade show
14:00 – 15:00 Question and Answer Session
Delegates will be asked to submit questions to a panel of experts. Questions can be submitted before the event or on the day
15:00 – 15:30 Dry Powder Mammalian Cell Culture media production: From Easy going to sophistication –
a case study DMEM / F12 production
Dr. Jörg von Hagen, PMP , Head of Process Development / Launch Management, Pharm Chemicals Solutions,Merck Millipore Division, Germany
This talk will discuss Dry Powder Mammalian Cell Culture media production-From Easy going to sophistication – a case study DMEM / F12 production. Points will include :
Physico chemical characterisation of DMEM F12
Batch to batch consistency: the grail in media production and supply
Impact on cellular performance of Chinese Hamster Ovary cells
Media testing beyond cell growth
Perspectives in future media preparation
15:30– 16:00 Afternoon Tea/Coffee and trade show
16:00 – 16:30 Recent advances in photoporation technology for cell injection and transfection
Dr Maria Torres, University of St Andrews, Scotland, UK
The injection of exogenous materials into mammalian cells is an important and ubiquitous procedure in cell biology. Recently, the use of laser was realized as an effective means to transiently permeabilise the cell membrane allowing intracellular delivery of a variety of sustances such as plasmid DNA, siRNA, mRNA or nanoparticles. This technique called photoporation is a gentle, robust and highly efficient method for cell transfection. In this presentation, the recent progress in this field and the development of the photoporation technology will be discussed with the end users in mind.
16:30 – 17:00 Remodelling of mRNA translation in the cold during mammalian cell bioprocessing
Professor Mark Smales, University of Kent¸UK
Reduced temperature cultivation of mammalian cells is often used duirng both stable and transient expression of recombinant proteins to improve the yield and product quality. Upon such cold shock, mammalian cells activate a number of cellular responses including remodelling of mRNA translation. Here we will describe our understanding of this remodelling and the implications for bioprocessing.
17:00 Chairman’s summing up
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About the Chair
John Davis is Visiting Lecturer in Biotechnology at the University of Hertfordshire, and Chairman of the UK Branch of the European Society for Animal Cell Technology (ESACT-UK). After a degree in Biochemistry at Sheffield, he moved in 1974 to Renato Dulbecco’s laboratory where he was initiated into the art of cell culture. Following PhD studies in Leicester, he moved to Switzerland, working with both Norman Iscove and Georges Köhler, the latter starting him on his many years of research into the use of monoclonal antibodies, particularly in therapy. After a further postdoctoral position, at the University of Cambridge where he worked on the early stages of the development of Campath (Alemtuzumab; now also known as Lemtrada), he made the transition to industry, working first for PA Technology and then (for nearly 20 years) for the Bio-Products Laboratory. In 2007 he made the transition back to academia. In addition to undergraduate and postgraduate teaching, he now runs open courses on Basic Cell Culture and Intermediate/Advanced Cell Culture. He has served on both the UKCCCR subcommittee on the Use of Cell Lines in Cancer Research, and the EC task force on Good Cell Culture Practice. In addition he has edited a number of books on cell culture, including Basic Cell Culture: A Practical Approach, and (with Glyn Stacey) Medicines from Animal Cell Culture. His most recent book is Animal Cell Culture: Essential Methods, which was published in March 2011 by Wiley-Blackwell
About the Speakers
Maria Torres is currently a postdoctoral research fellow in Biophotonics at the University of St. Andrews, Scotland. She did her PhD studies in the same university, having obtained a prestigious Scottish University Physics Alliance (SUPA) prize studentship for her PhD. She works in collaboration with biologists in the field of neuroscience, breast cancer and embryology among many others to use the novel technique optical transfection for advance biological applications. Her expertise is on using a variety of lasers and microscopy systems to perform optical manipulation of single cells and developing embryo.
Katy Petherick is in her final year of a PhD investigating the role of autophagy in colorectal cancer at the University of Bristol, supervised by Professor Chris Paraskeva in the Tumour Biology group. Prior to commencing her PhD, Katy completed her degree at the University of Birmingham, which involved a year in industry at Cambridge Antibody Technology (now MedImmune).
Gary Lye is Professor of Biochemical Engineering within the Advanced Centre for Biochemical Engineering at University College London (UCL). He is Deputy Head of Department and Director of the Industrial Doctoral Training Centre (IDTC) in Bioprocess Engineering Leadership. He received his PhD in Biotechnology from the University of Reading in 1992. Between 1993 and 1996 he was successively a Research Fellow and then Lecturer in Chemical Engineering at Imperial College London and the University of Edinburgh. He joinedUCL in 1996. He has broad research interests on the application of microscale and automation techniques to rapid bioprocess design, optimisation and scale-up.
Thorsten Adams, molecular biologist, is product manager fermentation technologies at Sartorius Stedim Biotech. He was educated at the University of California, Berkeley and the University of Goettingen, where he obtained his Ph.D. from the department of microbiology and genetics in 2005. He worked as a scientist in technology development in several positions, latest at Morphosys in Munich, Germany. He developed cell culture expression and screening systems and is experienced in cell culture development using several production systems to large scale bioreactors. Dr. Adams was amongst the first users of single use upstream technology in the industry. He authored several papers, book chapters and patents. In 2007, he joined Sartorius Stedim Biotech, where he is responsible for product management of single use bioreactors. He is an expert for the implementation of single use bioreactor technology in upstream processes.
Jörg von Hagen is head of Merck Millipore Process Development R&D in Darmstadt (Germany). Having studied biotechnology and signal transduction in Giessen and Darmstadt, he received his academic degree with an award-winning thesis in 2001. Dr. von Hagen has more than 20 years of practical expertise in biotechnology, especially in molecular cell biology and proteomics.
Dusko Ilic, obtained his MD degree and BSci in Molecular Biology at the University of Belgrade, PhD at the Tokyo University, Japan, and postdoctoral training at the University of California in San Francisco. Before joining King’s College School of Medicine in London as a Senior Lecturer in Stem Cell Science, he held positions of Adjunct Associate Professor at the University of California San Francisco, Consultant at the Veteran Affairs Medical Center, San Francisco, and the Director of R&D at StemLifeLine, a California-licensed stem cell company. His current research interest lies in human embryonic stem (hES) cells, induced pluripotent stem (iPS) cells, cancer stem cells, and regenerative medicine.
Mark Smales work focusses upon furthering our understanding the cellular processes underpinning recombinant protein synthesis and quality from mammalain cells. He is currently Professor of Biotechnology at the University of Kent and the Director of the Center for Molecular Processing at Kent.
Keywords: cell, transfection, lasers, microscopy, Autophagy, Puncta, p62, Flux, cell, transfection, lasers, microscopy, dry powder, mammalian cell culture media, batch to batch consistency, human embryonic stem cells, clinical grade, cGMP, bioreactor, process development, large scale, upstream processes, recombinant protein production; cold-shock; CHO cells; cell engineering
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Price of Attendance
The Penridge Suite
470 Bowes Road