Gene expression profiling of epithelial cells from human cervix cultured in Organ Chip and transwells

Modulation of mucus production by the human ecto- and endo-cervical epithelium by steroid hormones and associated interactions with commensal microbiome play a central role in the physiology and pathophysiology of the female reproductive tract. Here, we show that in a human organ-on-a-chip model of cervix (Cervix Chip), use of continuous fluid flow promotes ecto-cervical differentiation, whereas use of periodic flow including periods of stasis stimulated endo-cervical specialization. Continuous flow exhibited significant upregulation of genes associated with ectocervical epithelial phenotype whereas the same cells cultured in the same medium under periodic flow or under static conditions in Transwell inserts upregulated genes more closely associated with the endocervical epithelial phenotype. Similar results with minor differences were obtained using epithelial cells isolated from three donors each from a different ethnic background (African American, Hispanic, and Caucasian). Epithelial cells from three donors have been cultured in two experimental arrangements: a) organ chips at low strain cycle (n = 2 technical replicates) and b) medium strain cycle (n = 3 technical replicates) and c) static transwells (n = 3 technical replicates). Gene expression in the epithelial cells was measured using RNAseq.