cultivation_medium_for_chlamydophila_pecorum

Overview

External ontology

mediadive.medium:1503 (source link unavailable) — CULTIVATION MEDIUM FOR CHLAMYDOPHILA PECORUM

Applications

• Microbial cultivation

Notes

Source: DSMZ | Link: https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium1503.pdf

Composition

flowchart LR
    CultureMech_000973["cultivation_medium_for_chlamydophila_pecorum"]
    ing0_MEM_Medium["MEM-Medium"]
    CultureMech_000973 --> ing0_MEM_Medium
    ing1_Fetal_bovine_serum["Fetal bovine serum"]
    CultureMech_000973 --> ing1_Fetal_bovine_serum
    ing2_Aminoacids["Aminoacids"]
    CultureMech_000973 --> ing2_Aminoacids

Ingredients (3)

Preparation

1. Cultivation of CACO-2 cells (ACC 169)
2. Filter-sterilize (0.2 µm) and keep no longer than 4 weeks. Store at room temperature to facilitate detection of contamination.
3. Prepare a 25 cm2 flask and seed cells according to standard protocols (see DSMZ catalogue for ACC 169). Incubate at 37°C plus 5% CO2. When a confluent layer has formed, infection can be carried out.
4. Exchange medium with 6 ml of Infection Medium (as above with the addition of 1 µg/ml cycloheximide (final concentration)) and add 500 - 1000 µl of EB stock solution (thawed quickly to 37°C).
5. Centrifuge for 1 h onto the cell layer at 1600 rpm at 20°C.
6. Incubate at 37°C + 5% CO2. Control cells daily and look for inclusions. Not all Chlamydiae form well-visible inclusions, ultimately, immunofluorescence or in situ-hybridization techniques are necessary to visualize inclusions.

Target organisms

No target organisms are recorded.

Provenance

Merged from 1 source record.

• cultivation_medium_for_chlamydophila_pecorum (CultureMech:000973)