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Animal Cell Culture Methods
and Scale-up Strategies |
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This introductory course emphasizes cell culture and
scale-up theory, as well as an introduction to laboratory techniques utilized in animal cell culture. These include modern techniques used with mammalian cell culture from frozen stocks to inoculation and cultivation in bench top bioreactors. Lectures provide the background to understand the principles of cell culture and bioreactor design. Participants will gain experience with a variety of bioreactor systems, including stirred tank bioreactors, microcarrier and hollow fiber systems. The course covers mammalian and insect cell culture systems and is highly recommended for those who want to learn cell line propagation, characterization, and quality control, and be introduced to the complex issues of scale-up of cell culture from the research bench to larger reactors.
Approximately 50% of the time is spent in lecture and 50% is in the laboratory. |
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Lecture:
• Mammalian cells in culture
• Medium formulations and serum-free medium
• Primary and continuous cell lines
Hands-on Laboratory:
• Cell growth and subculturing methods
• Primary cell culture from chicken embryo
• Establishing a primary cell line
Lecture
• Cell line characterization for production of biologics
• Monoclonal antibody production
• Transformation of mammalian cells with SV40 DNA
Hands-on Laboratory:
• Mycoplasma detection
• Fusion of cells in monolayer
• Cell transformation with viral DNA via transfection assay
Lecture:
• Validation, cGMP and change control
• ELISA theory
• Design and operation of stirred-tank bioreactors for animal cell culture
Hands-on Laboratory:
• ELISA techniques
• Stirred tank reactor design and instrumentation, pH calibration,
reactor preparation and sterilization
• DO and pH probes, probe maintenance, and calibration
Lecture:
• Achieving precision with pH measurements and dissolved oxygen
• Perfusion techniques in stirred tank bioreactors
Hands-on Laboratory:
• Growth of anchorage-dependent cells on microcarriers in stirred tank reactors
• Growth of suspension cultures in bioreactors
• Medium addition to bioreactors, cell counts, viability, seeding reactors
 (adjourns
at noon)
Lecture:
• Insect cell culture
• Scale-up strategies for insect cell culture
Hands-on Laboratory:
• Insect cell counting
• Plaque assays
John Enders, Ph.D., Delmont Laboratories
Richard Frisque, Ph.D., Penn State, Department of Biochemistry and Molecular Biology
Vikram Gossain, Ph.D., New Brunswick Scientific, Co., Inc.
Robert Lumley-Sapanski, Penn State, Department of Food Science
Andrea Mastro, Ph.D., Penn State, Department of Biochemistry and Molecular Biology
Michael McMurtrie, Broadley James Corporation
Khushboo Nangia, Centocor, Inc.
Bill Scheuchenzuber, Ph.D., Penn State, Huck Institutes of the Life Sciences
Beatrice Sirakaya, M.S., Penn State, Department of Biochemistry and Molecular Biology
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