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Tuesday, August 23
7:30am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee
Stem Cells: Biopreservation for Therapy
8:25 Chairperson's Remarks
8:30 Bioprocessing and Cell Storage Methods for Human Progenitor and Stem Cells
Mary Pat Moyer, Ph.D., CEO and Chief Science Officer, INCELL Corporation LLC
Integrated strategies, combined with new reagents and other tools for GMP bioprocessing and storage of cells derived from various tissues, are important to bringing new cell and immunotherapy products to clinical applications. Stored cells from living and cadaveric donor tissues (e.g., adipose, bone marrow, blood, skin), either alone or in combination with other products, have unique needs and similar requirements. These will be reviewed with regard to time and temperature of storage and in the context of their therapeutic, diagnostic or research applications.
9:00 Serum Free Cryopreservation of Adult Stem Cells
Ram Devireddy, Ph.D., Associate Professor, Mechanical Engineering, Louisiana State University
Developing effective techniques for the cryopreservation of human adipose derived adult stem cells could increase the usefulness of these cells in tissue engineering and regenerative medicine. Unfortunately, the use of serum and a commonly used cryoprotectant chemical dimethyl sulfoxide (DMSO) during cryopreservation storage restricts the direct translation of adult stem cells to in vivo applications. Our results suggest that post-thaw cell viability, adipogenic and osteogenic differentiability can be maintained even when they are frozen in the absence of serum and DMSO.
9:30 Hematopoietic Stem Cell Growth and Cryopreservation: Removing Animal Proteins from the Culture and Optimizing the Freeze Curve for Long Term StorageLinda L. Kelley, Ph.D., Director, Connell O'Reilly Cell Manipulation Core Facility Dana Farber Cancer InstituteThe talk will focus on the use of human platelet lysate(PL) as growth media to replace fetal bovine serum (FBS) for growth of bone marrow-derived mesenchymal stem cells (MSC). Data demonstrate that MSC grown in PL have a shorter doubling time, remain phenotypically characteristic of MSC and maintain multilineage differentiation capability. Secondly, the talk will emphasize the importance of controlled-rate freezing and optimized freezing conditions for long-term storage and maximal recovery of hematopoietic stem cells.
10:00 Networking Coffee Break with Exhibit and Poster Viewing
CHO Cells: Biopreservation for Production
10:45 Genotoxic Assessment on CHO-Cells of Three Cryoprotectants Commonly Used for Human Oocyte Vitrification
Blandine Courbière, M.D., Ph.D., Medical Faculty, Laboratory of Environmental Mutagenesis and Biogenotoxicology, Université de la Méditerranée Aix-Marseille II
The aim of our work was to evaluate the possible genotoxic activity of three cryoprotectants extensively used in vitrification techniques: dimethyl sulfoxide, ethylene glycol, and propylene glycol. For this purpose, a Chinese Hamster Ovary cell line, commonly used in genetic toxicology, was used to assess i) the induction of DNA primary lesions by the comet assay and ii) the persistence of chromosomal damages (micronuclei) by the micronucleus assay.
11:15 Cryopreservation of High Density Recombinant CHO Cell Banks
James Moldenhauer, M.S., Senior Scientist, Manufacturing Science & Technology, Eli Lilly and Company
Seed train expansion of recombinant CHO cells from a Working Cell Bank (WCB) cryovial for eventual inoculation into a bioreactor remains a manually-intrusive process that is time consuming and labor intensive, often requiring several sub-cultivations in shake flasks of increasing volumes. The frequent handling of cell cultures is a potential source of contamination at each expansion step, as well as a possible cause of variability in the final cell inoculum used for bioreactor seeding. By increasing the viable cell density of the WCB, one could reduce or eliminate the need for shake flask expansion steps prior to inoculation of a bioreactor. Along with minimizing aseptic open culture manipulations, reduction or elimination of cell expansion steps could increase scheduling flexibility for pilot plants and/or commercial production facilities. We have demonstrated the feasibility of a novel method for cryopreservation of GS-CHO cells as a "pelletized cell bank" used as a high density cell inoculum for direct seeding of bioreactors.
11:45 Cryopreservation to Maintain Continuity of the CHO Cell LineYvonne A. Reid, Ph.D., Manager, Scientist, Cell Biology Program, American Type Culture CollectionChinese hamster ovary (CHO) cell line was derived from the ovary of the Chinese hamster in the 1960s. Since then it has become one of the most commonly used cell line for biological, medical and commercial applications including the production of therapeutic proteins. For these very important activities, cryopreservation of CHO cells ensures reproducibility and continuity of the cell line.
12:15 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own
Cryopreservation: Meeting the Demands of Scale-up and Production
1:55 Chairperson's Remarks
2:00 Cryopreservation and Long-Term Storage of Mammalian Cell Lines in 50 - 100 mL Cryobags
Rüdiger Heidemann, Ph.D., Group Leader, Cell Culture Development, Bayer HealthCare LLCThis presentation will discuss the cryopreservation of a recombinant mammalian cell line in cryobags suitable for GMP-type cell banks. It will highlight the challenges and differences compare to conventional banking procedures using cryo vials. In addition recovery data for the long-term storage in liquid nitrogen of these cryobags are presented.
2:30 Optimizing Mammalian Cell-Line Cryopreservation and Revival through Effective Detection and Depletion of Non-Viable Cells
Christopher Gregory, Ph.D., Professor, MRC Centre for Inflammation Research, University of Edinburgh; CSO, ImmunoSolv Ltd.
An often forgotten consequence of cryopreservation is the pro-apoptotic stress response caused by storage at low temperature. Thus, viability assessments of cells immediately following revival are often erroneous because they fail to take account of the rapid cell death responses (sometimes referred to as "delayed onset" cell death responses) that subsequently occur when the cells are cultured at physiological temperatures. In this presentation, the effective measurement of cell viability – including assessment of the commitment of apparently viable cells to apoptosis post-revival – will be considered. Furthermore, the beneficial effects of dead-cell removal on culture re-establishment, illustrated with antibody-producing cell lines and human embryonic stem cells, will be demonstrated.
3:00 Disposable Technology in Cell Banking and Expansion to Meet Demanding Process Timelines
Eric Becker, Ph.D., Associate Director, Mammalian Cell Culture Process, Boehringer Ingelheim Pharma GmbH & Co. KG
This presentation will provide an overview over current cell banking activities and topics such as long term storage and new strategies in cell expansion.
3:30 Networking Refreshment Break With Exhibit and Poster Viewing
4:15 INTERACTIVE PANEL: How will Mammalian Cell Culture Be Further Innovated?As mammalian cell culture moves forward meeting increased demands for ever higher titer, how will the task of culturing cells be innovated? What are the emerging technologies and approaches that will bring cell culture to new heights?Please join this interactive panel discussion as cell culture experts share their insights on how cell culture will be innovated to provide adequate capacity for the biologics industry.Moderator: Tiffany D. Rau, Ph.D., Global Technology & Technical Manager, Pall CorporationPanelists:Paula Meleady, Ph.D., Senior Research Scientist and Programme Leader, Proteomics Core Facility, National Institute for Cellular Biotechnology (NICB), Dublin City UniversityYung-Shyeng Tsao, Ph.D., Senior Principal Scientist, Cell Culture, MerckJames C. Warren, Ph.D., Principal Development Engineer, Vaccine Manufacturing Sciences and Commercialization, Merck & Co., Inc.Jianguo Yang, Ph.D., Principal Scientist, Commercial Cell Culture Development, Commercial Process Development, Biologic R&D, Genzyme
5:15 End of Conference