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Culture medium recipe

1 Media for maintenance of infected amoeba cultures
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trypticase Soy Broth with Yeast Extract (TSY) 1000 ml - -
2 OR
Peptone-yeast-glucose medium (PYG) 1000 ml - -
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trypticase soy broth 30 g 30 -
Yeast extract 10 g 10 -
Distilled water 1000 ml - -
1 Adjust pH to 7.3, autoclave at 121°C
Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Proteose peptone 20.000 g 20 -
Glucose 18.000 g 18 99.911
Yeast extract 2.000 g 2 -
Sodium citrate dihydrate 1.000 g 1 3.4
MgSO4 x 7 H2O 0.980 g 0.98 3.976
Na2HPO4 x 7 H2O 0.355 g 0.355 2.501
KH2PO4 0.340 g 0.34 2.498
Fe(NH4)2(SO4)2 x 6 H2O 0.020 g 0.02 -
Distilled water 1000.000 ml - -
1 Adjust pH to 6.5, autoclave at 110°C or filter-sterilize
1 Infected amoeba cultures can be maintained e.g. in 25 cm2 flasks (e.g. Nunc flasks, VWR, 734-2081, Thermo-Scientific, 156340) at 20-30°C in an incubator (no special requirements concerning CO2 or light).
1 Infected Acanthamoeba cultures don't have to be transferred to fresh culture flasks as frequently as usually done for mammalian cell cultures. Amoeba cultures can be maintained in the same flask for several weeks or months. However, if the cultures reach a high cell density, medium should be exchanged. For that purpose amoebae are detached from the surface of the flask by knocking at the culture flask (no trypsin required, amoebae are only weakly adherent compared to most mammalian cells) and the medium (containing the majority of cells) is then completely removed. Fresh medium is added and the few remaining attached amoebae will continue to grow.
2 Alternatively, cultures can also be passaged regularly (e.g. weekly) by transfer of a small aliquot of a densely grown culture (after cell detachment, as described above) to a fresh flask containing medium.
3 If cultures contain few attached cells, but high amounts of cell debris, medium should be exchanged without prior cell detachment. Cultures usually recover fast.
4 Depending on the host cell strain, the bacteria may cause host cell lysis. To maintain such cultures, bacteria have to be transferred by adding culture supernatant (approximately 0.5 ml) to fresh culture flasks containing uninfected host cells (approximately semi-confluent culture) each 7-10 days.
Solution occurs in 2 other media (first defined in Medium 1500).
1 Bacteria released from host cells can be stored in
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sucrose-Phosphate-Glutamate buffer 1000 ml - -
2 at -80°C for several months.
3 Infected cell cultures (supplemented with 10% DMSO) are best stored in liquid nitrogen.
Solution occurs in 2 other media (first defined in Medium 1500).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sucrose 75.00 g 75 219.106
KH2PO4 0.52 g 0.52 3.821
Na2HPO4 x 2 H2O 1.53 g 1.53 10.778
Glutamic acid 0.75 g 0.75 5.098
Distilled water 1000.00 ml - -


Last modified: 22.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.5
Equipment needed: Autoclave, Filter
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Associated strains

All strains for this medium: 6

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Trypticase soy broth 30 -
Proteose peptone 20 -
Glucose 18 99.911
Yeast extract 2 -
Sodium citrate dihydrate 1 3.4
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Cultivation metadata from 6 strains BacDive

phyla 2 6 strains
temperature mesophilic 5 strains