Division of Biology and Medicine
Center for Alternatives to Animals in Testing


The Boekelheide lab is using the prostate microtissue as a means to identify endocrine active chemicals that mimic or counteract the action of normal hormones.

Prostate 3D Microtissues

  • The Boekelheide lab has developed a 3D co-culture system of the human prostate. The co-culture microtissue comprises a mix of human prostate stromal cells and epithelial cells to mimic physiological stromal-epithelial interactions, which are critical in the development and normal function of the human prostate.
  • In this system, the epithelial-to-stromal cell ratio was fixed according to normal human physiology. The epithelial cells and stromal cells aggregate and spontaneously self-organize to form a microtissue with an epithelial compartment, a stromal compartment, and an active interface in between.
  • Immunostaining studies showed that cytokeratins 5/6 (CK5/6) were expressed evenly across the epithelial compartment, while CK18 keratin showed distinctive expression in certain regions of the microtissue, indicating cell differentiation during the microtissue maturation process.
  • To assess the microtissue response to hormones, the microtissues were treated with androgen (dihydrotestosterone), anti-androgen (flutamide), and combinations. Both the self-organization process and the morphology of the prostate microtissues were androgen-dependent, indicating that this prostate microtissue was androgen-responsive.
  • The Opera Phenix high-content imaging system was used to capture the 3D structure of the prostate microtissue and extracted 3D image features for further analysis. Selected 3D image features could distinguish androgen versus anti-androgen exposure of the prostate microtissue.
  • We aim to develop the current co-culture system to a rapid, economical and reliable screening platform for endocrine-disrupting chemicals. 

Learn More

Dent, M. P., Madnick, S. J., Hall, S., Vantangoli Policelli, M., Bars, C., Li, H., Amin, A., Carmichael, P. L., Martin, F. L., & Boekelheide, K. (2019). A human-derived prostate co-culture microtissue model using epithelial (RWPE-1) and stromal (WPMY-1) cell lines. Toxicology In Vitro, 60, 203-211. PMC6717042

Kabadi, P. K., Vantangoli, M. M., Rodd, A. L., Leary, E., Madnick, S. J., Morgan, J. R., Kane, A., & Boekelheide, K. (2015). Into the depths: Techniques for in vitro three-dimensional microtissue visualization. BioTechniques, 59(5), 279–286.  PMC4804457