Figure 1.

Evaluation of the cultivability of frozen biobanked fecal microbiota. (a) experimental workflow. Fresh stool was directly collected from healthy adult volunteers into pre-reduced 10% glycerol in phosphate buffered saline (PBS). Samples were transferred into anaerobic workstation immediately, homogenized to make a 20% (w/v) fecal slurry and aliquoted followed by filtering using sterile gauzes to remove large particles. The pre-processed stools were cultured directly in anaerobic workstation or frozen in −80°C for up to 52 weeks prior to culturing with or without fructo-oligosaccharide (FOS) in triplicates. (b) Principal component analysis (PCA) score plot of quantified protein groups. The log10-tranformed LFQ intensities of protein groups that were quantified in >50% samples were used for PCA analysis. Different symbol colors indicate the weeks of frozen in −80°C prior to culturing, and different shapes indicate the treatment groups for each culturing experiment. Baseline, uncultured microbiome samples; QC, quality controls for monitoring mass spectrometry measurement performance; V31, V33 and V34, origin donors of the microbiome samples.