FIG 3.

Essential culture components required for M. pneumoniae growth. (A) Identification of groups of compounds in CMRL that are required for supporting growth in vB13. Data show the percentage of protein biomass relative to vB13 of different medium versions, in which commercial CMRL has been substituted with a full home-made CMRL version (hm-CMRL) or variants lacking specific groups of CMRL compounds. Cultures were grown in a 96-well-plate format and processed after 96 h of culture. The plus (+) and minus (−) symbols represent the presence or absence, respectively, of the indicated group of compounds (see Table S2 for a detailed list). Data represent the mean ± standard error of 3 independent biological replicates. Significance relative to the vB13 formulation was assessed by two-sided independent t test (*, P < 0.05; **, P < 0.005). (B) Identification of minimal medium constituents (vB13m) required for supporting M. pneumoniae culture. Data show the percentage of protein biomass relative to vB13 as described in panel A. Components that are common among the tested medium formulations are not shown for clarity purposes. The detailed formulation of vB13m can be found in Table 1.