Events

Workshop: Enabling Innovations for Cell Therapy Manufacturing

Date: 15 Nov 2011 - 15 Nov 2011

Venue: SIMTech Training Room, TOWER Block, Level 3

Introduction
Regenerative therapy is changing the practice of medicine; it will revolutionise healthcare over the coming years. Cell therapy manufacturing will require the development of innovative technologies to enable the large scale processing of functional living cells with expected production volumes. The aim of this workshop is to understand the needs and challenges of cell manufacturing for clinical use, to initiate a focused R&D program, and identify the supply chains to control cost of goods. A cell manufacturing process consists of upstream and downstream operations. Upstream processes include all steps of cell culture - the acquisition of tissue, isolation of cells, creation of cell bank, and expansion of cells to therapeutic quantities. Downstream process include cell harvesting, volume reduction, product purification, formulation for biopreservation, final container filling, cryopreservation, storage, testing, logistics, end user handling and delivery.  A*STAR’s biomedical research institutes are focusing on the upstream processing and optimisation. However, the downstream processes will need to be addressed as they become a major source of manufacturing bottleneck, which present an opportunity to engineering solution providers. The Singapore Institute of Manufacturing Technology (SIMTech) is bridging the research institutes, manufacturers and funding authorities to facilitate a workshop to discuss and explore next generation technologies to advance cell therapy manufacturing in Singapore. 

Programme

9.00am

Registration

9.20am

Welcome Address

9.25am

Manufacturing regenerative medicines: “one to many” translational research by Prof. David Williams, Loughborough University

10.20am

Challenges of manufacturing cell therapy products by Mr Kim David Raineri, Lonza Bioscience Singapore Pte Ltd

11.00am

Refreshment

11.30am

Pluripotent stem cell processing in Bioreactors by Dr Steve Oh, Bioprocessing Technology Institute, A*STAR

12.00pm

Defined matrices for expanding human embryonic stem cells by Dr William Birch, Institute of Materials Research and Engineering, A*STAR

12.30pm

Roundtable Discussion

1.00pm

Lunch & Networking

2.00pm

Large scale GMP production of an encapsulated cell therapy product for the treatment of solid tumour by Dr Brian Salmons, SGAustria

2.40pm

Thehollow fiber bioreactor and its potential for manufacturing bloodby Dr Mayasari Lim, Nanyang Technological University, School of Chemical & Biomedical Engineering

3.10pm

Measurement and inspection for the in-vitro culture of stem cell by Dr Li Xiang, Singapore Institute of Manufacturing Technology, SIMTech, A*STAR

3.30pm

Further Discussion and Wrap Up

4.00pm

End

 

Abstracts 

Manufacturing regenerative medicines: “one to many” translational research
Regenerative medicines have the potential to make a significant difference to the lives of many people. Realising this difference requires research and innovation in both “bench to bedside” and “one to many” translational sciences. “One to many” translation particularly requires the solution of manufacturing and supply problems. Regulation and Good Manufacturing Practice – extreme levels of repeatability and reliability to ensure the product delivered is the same as the one that went through clinical trials – demand high precision process and measurement solutions. Processing living biological materials that ultimately become the product adds significant complexity. This presentation will show the results of work demonstrating the precision automated manufacture of regenerative medicines in particular the expansion of populations of human stem cells. Automated adherent culture of human mesenchymal stem cells, neural progenitor cells for therapeutic use and human embryonic stem cell lines will be discussed. Process transfer conditions for each automated culture process being the ability to culture a cell population equivalent to an equivalent specification as that manually cultured by the particular collaborator. It will also describe quality engineering techniques for precision process design and improvement. These include the measurement and comparison of process capability for the manual and automated culture of a human cell line and the use of screening experiments and response surface methods to establish process conditions.  The presentation will also identify the requirements for measurement systems evolution for such therapies recognising that the precision required in the characterisation of output of the cell culture process will increase with the demands of the regulator and other stakeholders. It should be recognised that characterisation challenges arise from both measuring in vitro analogues of in vivo functional performance and from the requirement to absolutely control contamination and cross contamination and other engineering issues. The presentation will close by over viewing some of the core issues that remain to be addressed at each level of the manufacturing and supply hierarchy and work in progress at Loughborough that is addressing some of these issues including three dimensional construct manufacturing, product transport conditions and regulatory science. The presenter would particularly wish to acknowledge contributions of his colleagues in the Healthcare Engineering Group and their collaborators to the work above and the financial support of the Engineering and Physical Sciences Research Council. 

Challenges of manufacturing cell therapy products
Kim Raineri will present on Lonza’s contract manufacturing experience with adult stem cells at their Walkersville Maryland site.  He will review the relevant challenges associated with the scale-up of cell manufacturing including cell culture, purification, fill-finish, and cost of goods.  He also will discuss Lonza’s current approach to the evaluation and development of cell processing technologies to address. 

Pluripotent stem cell processing in bioreactors
The ability of human pluripotent stem cells (hESC and hiPSC) to differentiate to a variety of cell types generates a unique potential for the development of new cell based therapeutics and human based in vitro drug screening and testing. One of the issues that need to be resolved in order to develop these technologies is large scale, suspension stem cell bioprocessing.  I will present our platform technology using different microcarriers for expansion of pluripotent stem cells and differentiation to human cardiomyocytes and neural stem cells. Cell yields are typically at least 3 times higher and more consistent when generated in the versatile microcarrier system compared to the 2D or embryoid body culture platforms; and the processes require much less manual interventions as the volumes increase. There is the added option of further improvement by optimisation and control of environmental parameters.

Defined matrices for expanding human embryonic stem cells

The standard for culturing human embryonic stem cells (hESC) has relied on a layer of supporting feeder cells or MatrigelÔ, an undefined substrate consisting of a basement membrane extract from a murine sarcoma. Artificial surfaces that present extracellular matrix proteins or peptide sequences containing an adhesion-promoting motif are shown to support hESC expansion. These defined polymer substrates, when used in conjunction with serum-free cell culture media, offer a superior environment for the culture stem cells to be used in therapeutic applications.  Tissue culture polystyrene (TCPS) substrates coated with vitronectin (VN) or Laminin (LN) support the long-term expansion of pluripotent hESC. TCPS coated with a VN surface density above a threshold of 250 ng/cm2 exhibits a performance indistinguishable from Matrigel, despite significant differences between these matrices for hESC propagation.  Our recent activity focuses on implementing this defined environment for hESC expansion in a highly scalable technology, which is suitable for use in bioreactors. Polystyrene microcarriers, coated with VN or LN, are a defined 3D culture substrate, which promotes hESC adhesion and is capable of supporting their long-term expansion in a defined, serum-free culture medium. 

Large scale GMP production of an encapsulated cell therapy product for the treatment solid tumour
SGAustria has developed an encapsulated cell therapy for the treatment of solid tumours that allows lower doses of chemotherapy to be used thus reducing side effects and toxicity while at the same time increasing local anti-tumour efficacy. The safety and efficacy of the encapsulated cell therapy has been successfully demonstrated in two clinical trials and these will be presented. In order to develop this further as a marketable product, a large scale GMP production was established. Some of the critical issues and hurdles in setting up this process will be presented.

The hollow fiber bioreactor and its potential for manufactuirng blood
Hematopoiesis is an essential process of the body responsible for the lifetime supply and regeneration of blood. In vitro, the study and ex vivo expansion of blood performed in tissue culture polystyrene (TCP) or blood bags are often hampered by the lack of resemblance of culture microenvironments to that of the in vivo system – the bone marrow. This compromises the quality and quantity of ex vivo expanded hematopoietic stem/progenitor cells (HSPCs), in vitro evaluation of leukemic cell therapy, and basic research in experimental hematology. Here, we evaluate the potential of a hollow fiber bioreactor (HFBR) as an artificial bone marrow model system for large-scale ex vivo expansion of hematopoietic cells. The unique advantages offered by the HFBR is its ability to achieve high cell densities that closely mimics the bone marrow stroma, provide a three-dimensional microenvironment for an in vivo-like cell-ECM and cell-stroma interaction, and enable perfusion which ensure continuous exchange of nutrients and waste materials. We have successfully established a hematopoietic co-culture within the HFBR using HS5 as the stroma support. The stromal culture was maintained for 28 days and was able to achieve good glucose consumptions of under 1g/L per day and produce a continuous supply of an array of hematopoietic cytokines. In the co-culture system, using K562 cells, the HFBR is able to expand over 80 times more cells than that achieved in TCP after 2 weeks. As with a standard co-culture, the HFBR was able to generate various myeloid lineages but did not appear to have tendencies toward specific lineages as observed in TCP. Moreover two distinct, adherent and loose, populations can be retrieved in the HFBR system. Our results demonstrate feasibility of the system and present exciting avenues to be explored in cell-stroma/ECM studies and clinically relevant ex vivo expansion or blood manufacture. 

Measurement and inspection for the in-vitro culture of Stem Cell 
A stem cell imaging System (SCIS) has been developed which provides non-invasive quantitative method of recording the growth of live hESC. Using imaging processing and proprietary algorithms, images of cell growth are captured daily and analysed to produce quantitative parameters such as seeding density and cluster size. These parameters, plotted with time scale, are valuable in determining the growth rate and quality of growth of the stem cell cultures. A dual field 3D imaging system will also be presented which is used for the assessment of 3D clusters cultured on microcarriers. With further development, this system could prove to be useful tool for hESC manufacturing process quality control. 

About the Speakers 

David Williams has been “Star Professor” in Healthcare Engineering at Loughborough University since 2003. He is Director of the university Research School of Health and Life Sciences. He leads national centres for research led innovation in regenerative Medicine in collaboration with Nottingham and Keele Universities and industry and agency partners, in particular the EPSRC Centre for Innovative Manufacturing in Regenerative Medicine. Until 2003 David was Technical Director of Bespak, a major supplier of drug delivery devices to the pharmaceutical industry. This followed ten years as Professor of Manufacturing Processes at Loughborough. His early career, while including industrial jobs with GKN and Metal Box, was primarily within the Cambridge University Engineering Department. David was elected a Fellow of the Royal Academy of Engineering in 2002 and holds the PhD degree of Cambridge University and DEng degree of UMIST/University of Manchester. David was an advisor to GINTIC-IMT from 1991-1997. 

Kim David Raineri is the Business Director for Lonza Bioscience Singapore Pte Ltd.  He is currently responsible for the start up and ongoing buisness operations of Lonza’s Cell Therapy contract manufacturing operation in Singapore.  Prior to this position, he was the Director of Operations for Lonza Walkersville (Maryland USA), where he managed multiple operations at the site including Cell Therapy contract manufacturing, manufacturing of  Endotoxin detection (LAL) testing kits, powdered and liquid cell culture media, and other research products marketed under brand names Biowhittaker™, Clonetics™, and Poietics™. Prior to Lonza, he was with CryoLife (Georgia USA) as the Senior Manager of the Tissue Processing Lab.  In this role, he was responsible for daily operations of human allograft manufacturing including human heart valves, vascular and orthopedic grafts.  Kim Raineri has a Master’s of Business Administration from Kennesaw State University and Bachelors of Science from University of Miami. 

Steve Oh received his PhD in Biochemical Engineering in Birmingham University, UK in 1990 and he is currently Principal Scientist and Associate Director at the Bioprocessing Technology Institute (BTI), one of the biomedical research institutes under the Agency for Science Technology and Research (A*STAR). Since 2001, together with Dr Andre Choo they have nucleated and grown the Stem Cell Group in BTI with particular focus on bioprocessing issues related to human embryonic stem cells (hESC) resulting in >40 published papers and 9 filed patents. The group’s main areas of research are stem cell characterisation, serum free, feeder free culture and scale up. Most recently, the group has developed a microcarrier platform technology on defined extracellular matrices which is amenable for bioprocess control and potential commercial scale production of pluripotent hESC, hiPSC, human mesenchymal stem cells, cardiomyocytes, neural stem cells and osteoblasts. His vision is to create viable bioprocesses for cell therapy, and drug development applications with allogeneic stem cells. 

William Birch received his PhD in Physics from Carnegie Mellon University and is currently Senior Scientist at the Institute of Materials Research and Engineering, A*STAR. Prior to moving to Singapore, he worked at Corning Inc., innovating of a product that rapidly became the market reference for immobilised DNA arrays. From a background in molecular adsorption and self-assembly on silica-based glass to immobilising bio-molecules on polymeric substrates, his current activities focus on engineering tunable bioactive surface properties. In collaboration with biologists, the bio-response of living systems to these defined cell culture substrates enables the development of materials applications for cell culture and tissue regeneration.  

Brian Salmons received his PhD in London and after research positions in the U.S.A., Switzerland, and at the Ludwig Maximilian University, Munich, Germany, he became the Scientific Director of the European biotech company, Bavarian Nordic, and was involved in taking the company to IPO. During this time, Dr Salmons co-invented a cell encapsulation technology shown to be safe and efficacious in a phase I/II clinical trial published in The Lancet. He co-founded the Biotech company, Austrianova, a spin out of the Veterinary Medicine University in Vienna, and was instrumental in obtaining orphan drug status from the European Medicines Agency for its lead product. In 2007, Dr Salmons co-founded and became the CEO of Austrianova Singapore Pte Ltd (SG Austria), an independent company established to further develop the encapsulation technology as products for a variety of indications such as various forms of cancer, diabetes, neurodegenerative, cardiovascular and infectious diseases. He is the author or co-author of over 120 peer reviewed scientific articles and inventor of 7 patent families. 
  
Mayasari Lim obtained her PhD degree in Chemical Engineering from Imperial College London and her B.Sc. degree in Chemical Engineering from the University of California at Berkeley. Prior to pursuing her graduate studies, she worked as a process and research development engineer in the semiconductor industry for 4 years and filed two US research patents during this time. She is now an assistant professor at the School of Chemical and Biomedical Engineering in Nanyang Technological University in Singapore. Her research focus is in stem cell bioprocess engineering and she has contributed toward the area of stem cell culture characterisation, bioprocess development, process optimisation and cell culture monitoring technologies. 

Li Xiang obtained his PhD in Tsing Hua University, Beijing. He worked in Aiwa Singapore Pte Ltd  for five years  as a senior engineer with expertise in mechanical design and optical data storage technology. After that he joined Dataplay Pte Ltd as a senior optical engineer, incharge of optical pickup system.  He has been working in SIMTech from Aug 2001. The main research areas include optical measurement, inspection, and bio-imaging technology. 

Who Should Attend
CTO, R&D managers, engineers and researchers, academic staff and students.

Registration
Pre-registration for this non-chargeable event is necessary for logistics and catering purposes. Seats are available on a first-come, first-served basis.
To reserve a place, please register online.

Contact Us
For technical enquiries, please contact Dr Li Xiang, Email: lixiang@SIMTech.a-star.edu.sg
For general enquiries, please contact Alice Koh, Email: kohth@scei.a-star.edu.sg