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Culture Media

The successful identification and longer term maintenance of amoebal strains requires that the culture be clonal and that all the nutritional requirements have been met to sustain the amoeba indefinitely.  Most amoebae are cultured monoaxenically, that is in the presence of a food organism, usually a bacteria, that is typically E. coli.  A few amoebal species have been axeniced, that is culture conditioned have been determined that sustain the amoeba without accompanying organisms so that they get all their nutrition from the broth.  This method of culture is beneficial for biochemical analysis and has allowed both Dictyostelium and Acanthamoeba to be developed into favourite model organism for cell biology research. 

Many different recipes are available (CCAP Catalogue of Strains 1995) so I will mention a few that are new or unusual.

 

YELP.  This is a recipe I used successfully to culture a group of marine amoebae.  This is based on the modified Chang's medium for Naegleria.  Make up the following ingredients in 75% seawater and autoclave for 20 minutes at 110oC.  Its often best to filter before use to remove precipitants.

Source  g/litre
Yeast extract Difco 0127-07-1 5
Liver digest Oxoid L27 3.2
Peptone Oxoid L37 11
Glucose Sigma  G-7528 10
Na2HPO4 Sigma  S-7907 0.51
KH2PO4 Sigma  P-0662 0.486
 

AX2. This is the medium of choice for the culture of Dictyostelium isolates (AX2, and NC4).  I have found this media suitable for all axenic strains of Acanthamoeba including some 200 strain I have isolated from soil.  Substitution (g for g) of tryptone for peptone increases cell density achieved with some strains.

Source  g/litre
Peptone Oxoid L37 14.3
Yeast extract Difco 0127-07-1 7.15
Glucose Sigma  G-7528 15.4
Na2HPO4 Sigma  S-7907 0.51
KH2PO4 Sigma  P-0662 0.486
 

Pollne-Fuller's axenic medium for Trichosphaerium

 PES medium (Provasoli Enriched Seawater
0.8% nutrient broth, 0.5% yeast extract, 0.2% sucrose
References

 

 

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