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

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