1. Serum-Free Media Chapter 10

2. 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

3. 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

4. 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

5. Disadvantages of Serum  Quality Control -Changing batch serum (replacement) requires extensive testing -Plating efficiency, Checking growth curve, Preservation of cell culture characteristics and Sterility

6. Disadvantages of Serum  Specificity -Several batches of serum should be held on reserve -Coculturing different cell types can be a problem

7. 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

8. Disadvantages of Serum  Downstream Processing -Presence of serum creates major obstacle to product purification -Limit the pharmaceutical acceptance of product

9. Disadvantages of Serum  Contamination -Viruses and Mycoplasma -Country of origin and batch number

10. Disadvantages of Serum  Standardization -Standardization of experimental and production protocols is difficult

11. 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

12. 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

13. 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

14. 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

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