RNA sequencing of HCl- and Lactate-treated ectocervical organoids

We sought to investigate the role of Lactobacillus in the self-renewal and differentiation of human cervical stem cells. Since the basal cells reside underneath of multiple layers of parabasal and suprabasal cells, we reasoned that Lactobacillus communicates with stem cells mainly using its metabolite rather than direct contact. L. crispatus is one of the most common Lactobacillus species in the human cervicovagina. Thus, we examined the effect of L. crispatus cell free supernatant (CFS) in the cervical organoid formation. Since the CFS of L. crispatus is more acidic than the uncultured control media (pH 7.07 vs 7.35), we adjusted to the same pH by adding hydrochloric acid (HCl) to the control media. Lactic acid (LA) was the top enriched metabolite in the L. crispatus CFS. We performed bulk RNA sequencing of HCl- and LA-treated ectocervical organoids to determine the global transcriptional response by LA treatment.