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Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
This preparation has tested positive for Mycoplasma contamination.
Note that activities with high potential for aerosol production require BSL-3 facilities and practices.
Forms intracellular inclusions which react with specific anti-C. pneumoniae monoclonal antibodies and monoclonal antibodies directed against the chlamydia LPS.
Inclusions will not stain with iodine or monoclonal antibodies against the MOMP of C. trachomatis. Infection is enhanced by centrifugation and use of cycloheximide (1 to 2 µg/mL) in medium.
Suggested protocol for propagation: Add glass beads and vortex preparation to disrupt cells. Infect monolayer with disrupted material. Centrifuge at 3000 x rpm (750 x g) for 1 hour.
Feed with fresh growth medium containing FBS prescreened for Chlamydia antibodies and 1-2 µg/mL cycloheximide. Incubate at 37°C for 72 hours.
Infection is enhanced by centrifugation and use of cycloheximide (1 to 2 µg/mL) in medium.
Incubation: 2-3 days at 35°C to 37°C in a 5% CO2 atmosphere in air.
Recommendations for Infection: For best results cells should be 24 to 48 hours old and 80%-95% confluent.Chirgwin K, et al. In vitro susceptibilities of Chlamydia pneumoniae (Chlamydia sp. strain TWAR). Antimicrob. Agents Chemother. 33: 1634-1635, 1989. PubMed: 2817862
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