Preparation of Artificial Sputum Medium to Emulate Cystic Fibrosis Lung Sputum Ex Vivo

Cystic fibrosis (CF) is a genetic disorder that causes thick, sticky mucus accumulation in the lungs, forming sputum that supports pathogenic microbial growth.

To prepare artificial CF sputum, take solutions with key biochemical properties.

The mucin mix contains mucins, which are high-molecular-weight glycoproteins, to replicate CF sputum viscosity and promote microbial adhesion.

The chemical mix supplies ions and glucose to support microbial metabolism.

The biological mix provides nutrients for microbial growth.

Combine all to form the artificial sputum medium (ASM).

Add a buffering agent to maintain a mildly acidic pH that favors pathogen survival.

For medium sterilization, transfer the ASM to a vacuum filtration chamber with a membrane filter.

Filter under agitation to avoid mucin aggregation and prevent filter clogging.

The vacuum draws the medium through the filter, removing microbial contaminants and macromolecular aggregates.

The ASM is ready for culturing CF-associated microbes.

Prepare an artificial sputum medium by adding 250 milliliters of ASCM to the 1 litre bottle containing ASMM. Then, add 250 milliliters of ASBM to the medium bottle. Titrate the medium with a basic 1 molar MOP