Day 1 | Day 2
Tuesday, August 25, 2009
7:30am Morning Coffee (Sponsorship Opportunity Available)
8:25 Chairperson’s Remarks
Edward Lee, Ph.D., President, BioPharm Consulting
8:30 The Impact of Cellular Cross-Contaminated or Misidentified Cell Lines
Yvonne Reid, Ph.D., Collection/Research Scientist, Cell Biology, American Type Culture Collection
Over the years numerous cell lines have been shown to be misidentified, due in part to lack of adequate characterization protocols available in previous years. Recent technological advances have given rise to an improved capability for cell line characterization. The financial loss incurred by misidentification and erroneous tools is estimated in the millions of dollars. An overview of the current technologies used to authenticate and characterize animal cell lines will be presented.
9:00 Physical Cell Measurements and Analytical Models for Quality Control and Assurance in Expanding Cell Cultures
John T. Elliott, Ph.D., Research Scientist, Biochemical Sciences Division, Cell Systems Sciences Group, National Institute of Standards and Technology (NIST/CSTL)
Large scale expansion of cell cultures is required to produce cells for frozen stock supplies and cultures that may be used in drug or toxin screening and medical therapy applications. Ideally, the cells at the end of the expansion phase are identical to the seed cells, but it is possible the culture characteristics are significantly modified by spontaneous phenotype switching, senescence and inadvertent cell contamination. We are testing the use of physical measurements such as cell volume and cell spread morphology in conjunction with analytical models to identify changes that may occur during expansion. The results of this study suggest that these tools will be useful to evaluate quality control and assurance in expanding cell cultures.
9:30 Biopreservation Considerations for Cell Culture - From Source Material to Finished Product
Aby J. Mathew, Ph.D., Senior Director, Strategic Relations and Senior Scientist, BioLife Solutions, Inc.
One aspect of bioprocessing development for cell culture, cell therapy, and tissue engineering that remains a bottleneck, is biopreservation of the source material and finished product. This presentation will address the science behind current biopreservation, the pitfalls that are both recognized and often unrecognized, and new preservation technologies that improve the final cellular product in terms of Efficacy, Quality/Regulatory Footprint, and Cost Efficiency.
10:00 Sponsored Presentation (Opportunity Available – Please contact Suzanne Carroll at scarroll@healthtech.com)
10:15 Networking Coffee Break with Exhibit and Poster Viewing
11:00 The XD Process: Development of a High Titer Process for PER.C6 Cells
Kathryn Golden, M.Eng., Associate Scientist III, Upstream Process Development, Percivia LLC
The latest advancements for the XD™ process, a high titer process using PER.C6® cells, aimed at maximizing the productivity of the process while minimizing the volume of medium consumption will be presented. This optimization strategy enabled the reduction of the total medium requirements by more than 50% from 40 bioreactor volumes to 15 bioreactor volumes while maintaining the maximum cell density at over 150 million viable cells/mL. In addition, an IgG antibody concentration of over 40 g/L of cell-free supernatant was achieved, which translated to over 27 g/L of harvest when corrected for biomass content produced in 17 days of culture time.
11:30 CHO-easyC - A Unique CHO Cell Line for the Development of Next-Generation Bioprocesses
Ferruccio Messi, Ph.D., Founder, President & Chief Executive Officer, Cell Culture Technologies, LLC
Recently, we derived a number serum-free cell lines such as NS0, Sp2/0, P3X63, BHK-21, HEK-293, Vero and COS from their respective serum-dependent parent cell lines, and adapted them to proliferation in minimal culture media. The serum-free cell lines were authenticated, certified and banked by the European Collection of Animal Cell Cultures (ECACC), and are currently available to the scientific community. Such cell lines can be routinely cultivated and banked in minimal culture media (MCM) totally free of proteins, peptones, peptides, chemically undefined additives and animal-derived ingredients. The MCM exclusively consist of small molecules identified by their respective CAS/EINECS numbers, and can be used for producing recombinant proteins, monoclonal antibodies and viral particles. Among the several biotech companies using the above serum-free cell lines to develop industrial bioprocesses, 15 companies already performed remarkable work on the cultivation of CHO cells in MCM. In particular, the CHO-K1 derived CHO-easyC cell line is currently being considered as one of the most promising host cell lines for the development of next-generation bioprocesses. Unique features and concrete applications of the CHO-easyC cell line will be presented.
12:00pm Using Systematic Metabolite Analysis to Guide Cell Culture Process Optimization
Zhaohui Geng, Ph.D., Principal Scientist, Culture Process Development, Pfizer, Inc.
During process optimization, our data indicated that elevated pH increased lactate accumulation in 1L bioreactor cultures of CHO cells. To investigate the molecular mechanisms of how elevated pH affects cellular energy production pathways, samples from elevated pH and control bioreactors were analyzed for intracellular and extracellular metabolites.
Metabolic analysis data indicated that many energy producing related metabolites in the pH elevated culture diverged dramatically from the control, particularly lipid metabolites. These changes appeared to center around more rapid consumption of glucose at day 3-5, followed by a metabolic switch to the use of amino acids for energy. After amino acids were depleted around day 9, triacylglycerols (TAGs) and phospholipids were hydrolyzed to supply energy needs with fatty acids. Metabolic analysis demonstrated that Glycerophosphorocholine (GPC), which is known to be liberated from membranes for combating osmotic stress as an osmolyte, was increased in the pH elevated culture compared to control.
The increased GPC suggests that osmolality may also have an important role in the metabolic switch. In other experiments, results suggest that high osmolality alone without elevating pH changed the glucose and amino acid metabolism. Therefore, osmolality might be an important factor to monitor during the pH optimization phase. The potential changes on lipid metabolism are currently under investigation. These results illustrate that systematic metabolite analysis could be used as a powerful tool to facilitate a rational approach to cell culture process optimization.
12:30 Lunch on Your Own (Lunch Presentation Opportunity Available)
1:55 Chairperson’s Remarks
Yubing Xie, Ph.D., Assistant Professor, The College of Nanoscale Science and Engineering, SUNY Albany
2:00 Opportunities to Improve Throughput and Timelines During Cell Line and Bioreactor Process Development to Decrease Time to Clinic
Alison Ridley, Ph.D., Senior Research Scientist, MedImmune
The talk will focus on areas where the cell line and bioreactor process development can be streamlined to reduce timelines. The talk also looks at implementation of new techniques to increase throughput and to allow for a more effective method to select manufacturing cell lines.
2:30 Application of Modern Design of Experiments for the Characterization of Mass Transfer in Bioreactors
Semsi Ensari, Ph.D., Group Leader, Senior Bioprocess Engineer, Fermentation and Scale-up, Ambrx, Inc.
To meet ever increasing needs for therapeutic proteins, it is essential to design processes that are cost-effective and robust. Modern design of experiments offers highly efficient solutions to characterize vital factors affecting the transport of nutrients to the cells producing lead candidate proteins. Classical approaches that vary one factor at a time are not effective when screening multiple factors and may be limited when coupled with theoretical correlations. Therefore, our current work is focused on the use of modern statistical designs to understand the interactions between variables and to optimize the mass transfer of gases. Screening experiments indicated that only few factors were critical to characterize mass transfer in the bioreactors. Selected factors were confirmed in subsequent experiments performed in larger bioreactors.
3:00 Exploring Small Scale Parallel Fermentor | Bioreactors Systems for Fast Track Development Sponsored by 
Christina Harper, MBA, Product Manager Re-Usable Fermentors / Bioreactors North America, Sartorius Stedim Biotech
A critical driving force behind research in bioprocess science and engineering continues to be the demand for fast and accurate analytical information that can be used, for example, to evaluate the interactions between biological systems and bioprocess operations. The ability to carry out large numbers of experiments rapidly and effectively is a known significant challenge. Developing efficient and practical bioprocesses frequently involves testing a large number of different strains and environmental conditions in various combinations at a small scale. However, the usefulness of these systems for selecting an organism that will be optimal under actual bioprocess conditions is limited. A discussion of the following points will provide example of how sophisticated small scale parallel screening fermentor | bioreactor systems can positively impact process developments and optimizations.
- increased process parameter measurements and control which allow an improved process understanding
- increased throughput allowing evaluation of more process impacts for finding superior process conditions
- consuming less resources like time and space so that resources for process developments can be increased
- used of stirred tank reactor design allowing for up- and down scalability
3:30 Networking Refreshment Break with Exhibit and Poster Viewing
4:00 Nanoengineered Systems for Cell Engineering and 3-D Culture
Yubing Xie, Ph.D., Assistant Professor, The College of Nanoscale Science and Engineering, SUNY Albany
The convergence of nanotechnology and cell biology has led to emerging approaches to deliver bioactive molecules to cells and integrate cells into micro/nanofabricated devices. These nanoengineered systems present exciting opportunities to serve as vehicles for therapeutic and biopharmaceutical applications. In this talk, the microfluidic synthesis of nanoparticles will be presented to deliver genes to cells and a 3-D cellular microenvironment will be exhibited to better capture the complex tissue physiology. The therapeutic potential of these cell culture technologies will be demonstrated.
4:30 The Animal Free Culture of Human Limbal Stem Cells for Transplantation
Sajjad Ahmad, M.D., Ph.D., Academic Clinical Lecturer, North East England Stem Cell Institute, Newcastle University
Limbal stem cells are required to regenerate the surface of the eye and are essential for normal vision. Limbal stem cell deficiency is a painful and blinding disease. One of the newer treatment options for limbal stem cell deficiency is the culture expansion and then transplantation of limbal stem cells. This technique is hampered by the requirement of animal cells or products within the culture. We have developed a novel animal free culture technique for human limbal stem cells which has enabled safe transplantation.
5:00 A Scalable Human Adipocyte Model for the Study of Insulin Resistance
Deb Nathan, Ph.D., Associate Research Fellow, Target Generation Unit, Biotherapeutics and Bioinnovation Center, Pfizer, Inc.
This presentation will describe the production of insulin responsive and resistant human adipocytes from bone marrow mesenchymal stem cells at scales ranging from 96-well plates to 2-4 million adipocytes in a T-175 flask. In insulin-sensitive cultures, insulin addition inhibits lipolysis (IC50 ~ 15 pM), stimulates phosphorylation of insulin receptor, IRS-1 and AKT (EC50 ~ 5 nM), and stimulates GLUT4 dependent glucose uptake (EC50 yields cells with mechanistically distinct insulin resistant phenotypes. ~ 0.5 nM). Treatment of insulin-sensitive cultures with TNF-alpha, insulin, or IL-1beta.
5:30 End of Optimizing Cell Culture Technology and Specialized Protein Expression Systems Conferences
For more information, please contact:
Mary Ruberry, Conference Director
Cambridge Healthtech Institute
Phone: 781-972-5421
E-mail:
mruberry@healthtech.comFor sponsorship information, please contact:
Suzanne Carroll, Manager, Business Development
Cambridge Healthtech Institute
Phone: 781-972- 5452
E-mail: scarroll@healthtech.com