Fig. 2

Culture of L. crispatus consortia in the vagina chip. A Total CFU/chip determined by quantifying non-adherent bacteria in effluents from the apical epithelial channel combined with counts of adherent bacteria measured within epithelial digests at 72 h compared to the original inoculum (T0). B Percent of OC1, OC2, OC3, and C0006A1 consortia bacteria adherent to the epithelium in vagina chips calculated by quantifying the bacteria recovered from the chip (digest) 72 h post-inoculation compared to the T0 inoculum. Significance was calculated by one-way ANOVA; *, P < 0.05 vs. C0006A1, ^#, P < 0.05 vs. OC1. C The pH values measured in the medium within the apical channel of the vagina chip cultured in the absence or presence of the OC1, OC2, OC3, and C0006A1 consortia at 72 h post-inoculation compared to pH measured in the basal channel. D Percent viability of vaginal epithelial cells assessed by calculating the number of live cells relative to control using trypan blue exclusion assay. In A–D, each data point indicates one chip; different colored points indicate chips from different donors; data are presented as mean ± s.d. In the lower graphs, results of metagenomics-based strain ratio analyses of the adherent bacteria from the L. crispatus OC1 (E), OC2 (F), and OC3 (G) consortia present within epithelial digests relative to the original inoculum are shown after 72 h of direct culture with the vaginal epithelium on-chip. 1–4 indicate results from 4 different chips