scgyem_medium

Overview

External ontology

mediadive.medium:1512 (source link unavailable) — SCGYEM- MEDIUM

Applications

• Microbial cultivation

Notes

Source: DSMZ | Link: https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium1512.pdf

Composition

flowchart LR
    CultureMech_000984["scgyem_medium"]
    ing0_Fetal_bovine_serum["Fetal bovine serum"]
    CultureMech_000984 --> ing0_Fetal_bovine_serum
    ing1_Casein["Casein"]
    CultureMech_000984 --> ing1_Casein
    ing2_Na2HPO4_x_7_H2O["Na2HPO4 x 7 H2O"]
    CultureMech_000984 --> ing2_Na2HPO4_x_7_H2O
    CHEBI_34683["CHEBI:34683"]
    ing2_Na2HPO4_x_7_H2O -.-> CHEBI_34683
    ing3_Glucose["Glucose"]
    CultureMech_000984 --> ing3_Glucose
    CHEBI_17234["CHEBI:17234"]
    ing3_Glucose -.-> CHEBI_17234
    ing4_KH2PO4["KH2PO4"]
    CultureMech_000984 --> ing4_KH2PO4
    CHEBI_63036["CHEBI:63036"]
    ing4_KH2PO4 -.-> CHEBI_63036
    ing5_Yeast_extract["Yeast extract"]
    CultureMech_000984 --> ing5_Yeast_extract
    ing6_Sucrose["Sucrose"]
    CultureMech_000984 --> ing6_Sucrose
    CHEBI_17992["CHEBI:17992"]
    ing6_Sucrose -.-> CHEBI_17992
    ing7_Na2HPO4_x_2_H2O["Na2HPO4 x 2 H2O"]
    CultureMech_000984 --> ing7_Na2HPO4_x_2_H2O
    CHEBI_91258["CHEBI:91258"]
    ing7_Na2HPO4_x_2_H2O -.-> CHEBI_91258
    ing8_Glutamic_acid["Glutamic acid"]
    CultureMech_000984 --> ing8_Glutamic_acid
    CHEBI_18237["CHEBI:18237"]
    ing8_Glutamic_acid -.-> CHEBI_18237

Ingredients (9)

Preparation

1. Medium for maintenance of infected amoeba cultures
2. Combine Solutions A, B and C and add
3. Maintenance conditions:
4. 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).
5. Maintenance/Passage of infected amoeba cultures:
6. 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. 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. If cultures contain few attached cells, but high amounts of cell debris, medium should be exchanged without prior cell detachment. Cultures usually recover fast. 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.
7. Adjust pH to > 8.0, autoclave at 110°C
8. Filter-sterilize
9. Filter-sterilize
10. Bacteria released from host cells can be stored in
11. at -80°C for several months.
12. Infected cell cultures (supplemented with 10% DMSO) are best stored in liquid nitrogen.

Target organisms

No target organisms are recorded.

Provenance