Transcript Serum-Free Media

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
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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
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