Getting started! I have ordered, received and stocked 3 strains at the DSM strain library. Unfortunately they do not have as many different options for vaginal bacteria, compared to all the isolates from the Human Microbiome Project for instances, and the strains I ordered were not all isolated from the vagina. I hope that they will display the same characteristics and can be used as a reference and to assist in setting up my methods. In the future I hope to with more vaginal isolates. I have
- Lactobacillus crispatus DSM 20584 (isolated from the eye)
- Lactobacillus iners DSM 13335 (isolated from human urine)
- Gardnerella vaginalis DSM 4944 (isolated from vaginal secretions)
To grow the strains I used MRS medium (liquid and plates) for L. crispatus and NYCIII medium for G. vaginalis and L. iners. Unfortunately I do not have access to an anaerobic chamber so I have been using an anaerobic jar. After closing the jar I use 3x: pulling a vacuum (minimally 0.8 bar) and filling with N2+CO2 gas. I grew plates and tubes over the weekend (72 hours) at 37 degrees and stocked them using 0.5 mL 60% glycerol + 1 mL of culture and stored them at -80 degrees. I am not sure how much the oxygen will affect the bacteria during handling and storage, especially Gardnerella is sensitive to oxygen. I will check later if the stocks lead to good growth. I will upload protocols for the media, the stocking and the anaerobic jar soon.