A simplified culture medium for an anti-inflammatory soluble factor production by L. reuteri CRL 1098

GRIET M, LANZA L, CAZORLA SI, GEREZ CL, FONT DE VALDEZ G, RODRÍGUEZ AV.

Mar del Plata

Congreso; LI Reunión Anual de la SAIB; 2015

Mundo Sano

Some specific soluble factors secreted by probioticlactic acid bacteria are able to modulate the immune response. Our previousstudies demonstrated that a Lactobacillusreuteri CRL 1098 soluble factor identified as a 5785 Da peptide, present inthe supernatant (LrS), modulates the inflammatory response triggered by LPS in vitro and in a mice model. LrS was producedunder stress conditions by starved cells after 4h incubation in RPMI 1640medium, where the bacteria maintained alive but did not proliferate. The aim ofthis work was to improve LrS production. Two steps will be performed: 1) Designof a simplified and low cost medium for optimal LrS production. Casitonemedium, PBS, TRIS, citrate buffer with and without glucose were assayed. Bacteriasurvival was evaluated by cell counting in MRS agar. LrS cytotoxicity wasdetermined by Trypan blue and MTT assay. L.reuteri only maintained alive in casitone medium and citrate buffer with 4g/l glucose. These LrS diminished TNF-αand nitric oxide production in both mouse peritoneal macrophages and RAW 264.7 cells stimulated with LPS. 2) Optimizationof medium components (MRS) for L. reuteribiomass production, using a factorialstatistical experimental model, is in progress. Application of an economic andmore competitive medium for LrS production by L. reuteri CRL 1098 represents an important step for itsenforcement at pilot scale.