Protocol Online logo
Top : New Forum Archives (2009-): : Microbiology

mycoplasma growth - (Apr/25/2009 )

Is it possible for mycoplasma to contaminate/grow in an LB culture of E coli?

-Lance-

Lance on Apr 25 2009, 05:51 PM said:

Is it possible for mycoplasma to contaminate/grow in an LB culture of E coli?

It is possible, though unlikely.
Although they generally have exacting nutritional requirements, there have been cases of mycoplasma/acholoplasma growing in simple microbiological media like LB or tryptose phosphate broth. These have usually occurred during validation of sterile production lines in the pharmaceutical industry.

-klinmed-

Mycoplasma growth factors in bovine serum fraction were separated by Sephadex G150 column chromatography and density ultracentrifugation. The major growth factor of bovine serum fraction eluted from the Sephadex column in the void volume. Its growth-supporting activity was greatly enhanced by the presence of bovine serum albumin in the mycoplasma culture media. Other investigators had previously identified the major growth factor in serum as an alpha-lipoprotein. Although density ultracentrifugation revealed the presence of traces of a high-density lipoprotein in bovine serum fraction, another, less dense component, isolated by ultracentrifugation (component 3) and containing cholesterol, cholesteryl esters, free fatty acids, triglycerides, and protein, but no lipoprotein, exhibited considerably more growth-supporting activity than did the high-density lipoprotein, thus indicating that at least two mycoplasma species do not require intact serum lipoprotein for growth. Both the high-density lipoprotein and component 3 exhibited maximum activity only in the presence of bovine serum albumin. A chloroform extract containing component 3 lipids combined with bovine serum albumin to form an effective, partially defined, less complex substitute for serum in mycoplasma culture media.A serological test is described which is designated the growth precipitation test and is based on the release of diffusible compounds from mycoplasma strains. During growth of mycoplasmas on solid substrates, precipitation lines are formed with antibodies from homologous antisera independent of heat-labile serum factors. The application of the test as a serodiagnostic tool was investigated by performing a complete serological cross-test with 10 mycoplasma strains.

-alexades-

A great tool to use for cell culture mycoplasma detection is the PCR mycoplasma detection kit offered by mdbioproducts.

Check it out:

mdbioproducts

Also, there tech-service department has been able to help me with multiple questions.

-researcher4life-