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Serum-Free Media Chapter 10 Change from serum to serum free media Serum can be reduced or omitted without apparent cell selection - if appropriate nutritional and hormonal modifications were made to media Disadvantages of Serum Physiological Variability – - Effects of albumin and transferrin are known - Effects of nutrients (amino acids, nucleosides, sugars, etc), peptide growth factors, hormones, minerals and lipids on cell culture is unknown Disadvantages of Serum Shelf Life and Consistency – - Serum varies from batch to batch - Might not last longer than a year - Should be replaced – might not be identical to first batch Disadvantages of Serum Quality Control - Changing batch serum (replacement) requires extensive testing - Plating efficiency, Checking growth curve, Preservation of cell culture characteristics and Sterility Disadvantages of Serum Specificity - Several batches of serum should be held on reserve - Coculturing different cell types can be a problem Disadvantages of Serum - Availability of serum Demand is increasing and can exceed supply Switch to serum-free media Average research laboratory uses 100-200L of serum per year - Commercial labs use more than 200L in a week Disadvantages of Serum Downstream Processing - Presence of serum creates major obstacle to product purification - Limit the pharmaceutical acceptance of product Disadvantages of Serum Contamination - Viruses and Mycoplasma - Country of origin and batch number Disadvantages of Serum Standardization - Standardization of experimental and production protocols is difficult Advantages of serum-free media Once practiced then becomes a selective media for that cell line (tables 10.1 and 10.2) Switch from growth enhancing medium for propagation to a differentiating inducing medium Disadvantages of serum-free media Multiplicity of media – Advantage to labs maintaining specific cell types but difficult for maintenance of many Selectivity – Some media may select a sublineage - Cells require different formulations Disadvantages of serum-free media Reagent purity – Degree of purity in reagents, water and cleanliness of apparatus needs to be high Cell proliferation – Growth is slower in serum free media Availability – of properly qualified-controlled serum-free media is limited - Products cultured are expensive Adaptation to Serum-Free media Carried over several serial subcultures Stable cell proliferation is established at one concentration - Subculture cells into a lower concentration until stable growth is reestablished - Dilute serum again This project is funded by a grant awarded under the President’s Community Based Job Training Grant as implemented by the U.S. Department of Labor’s Employment and Training Administration (CB-15-162-06-60). NCC is an equal opportunity employer and does not discriminate on the following basis: against any individual in the United States, on the basis of race, color, religion, sex, national origin, age disability, political affiliation or belief; and against any beneficiary of programs financially assisted under Title I of the Workforce Investment Act of 1998 (WIA), on the basis of the beneficiary’s citizenship/status as a lawfully admitted immigrant authorized to work in the United States, or his or her participation in any WIA Title I-financially assisted program or activity. Disclaimer This workforce solution was funded by a grant awarded under the President’s Community-Based Job Training Grants as implemented by the U.S. Department of Labor’s Employment and Training Administration. The solution was created by the grantee and does not necessarily reflect the official position of the U.S. Department of Labor. The Department of Labor makes no guarantees, warranties, or assurances of any kind, express or implied, with respect to such information, including any information on linked sites and including, but not limited to, accuracy of the information or its completeness, timeliness, usefulness, adequacy, continued availability, or ownership. This solution is copyrighted by the institution that created it. Internal use by an organization and/or personal use by an individual for non-commercial purposes is permissible. All other uses require the prior authorization of the copyright owner.