Generation and Characterization of an In Vitro Organotypic Foreskin Model for Future Study of the Penile Microbiome
Abstract
The foreskin is a site of HIV-1 acquisition in heterosexual males. The lack of relevant in vitro models that mimic the foreskin microenvironment, including innate immune and barrier functions, has limited our understanding of susceptibility at this site. We hypothesize that we can establish organotypic in vitro foreskin that mimics in vivo tissue. Organotypic foreskin models were generated and are composed of stratified cell layers that express E-cadherin suprabasally, filaggrin apically, and expressed TLR1, TLR2, and β-defensin-1 mRNA similar to in vivo foreskin. Preliminary results suggest organotypic foreskin tissues retain the ability to respond to LPS. Lastly, organotypic foreskin permeability appears high when subject to mechanical agitation. This work has established culture techniques to generate multi-layer epithelia and the methods necessary to characterize innate immune and barrier functions. This sets the foundation for future work to fully develop the model, including integrating immune cells, the microbiota, or HIV-1.