Three-dimensional podocyte-endothelial cell co-cultures: Assembly, validation, and application to drug testing and intercellular signaling studies

Min Li, Alessandro Corbelli, Shojiro Watanabe, Silvia Armelloni, Masami Ikehata, Valentina Parazzi, Chiara Pignatari, Laura Giardino, Deborah Mattinzoli, Lorenza Lazzari, Aldamaria Puliti, Francesco Cellesi, Cristina Zennaro, Piergiorgio Messa, Maria Pia Rastaldi

Research output: Contribution to journalArticle

Abstract

Proteinuria is a common symptom of glomerular diseases and is due to leakage of proteins from the glomerular filtration barrier, a three-layer structure composed by two post-mitotic highly specialized and interdependent cell populations, i.e. glomerular endothelial cells and podocytes, and the basement membrane in between. Despite enormous progresses made in the last years, pathogenesis of proteinuria remains to be completely uncovered. Studies in the field could largely benefit from an in vitro model of the glomerular filter, but such a system has proved difficult to realize. Here we describe a method to obtain and utilize a three-dimensional podocyte-endothelial co-culture which can be largely adopted by the scientific community because it does not rely on special instruments nor on the synthesis of devoted biomaterials. The device is composed by a porous membrane coated on both sides with type IV collagen. Adhesion of podocytes on the upper side of the membrane has to be preceded by VEGF-induced maturation of endothelial cells on the lower side. The co-culture can be assembled with podocyte cell lines as well as with primary podocytes, extending the use to cells derived from transgenic mice. An albumin permeability assay has been extensively validated and applied as functional readout, enabling rapid drug testing. Additionally, the bottom of the well can be populated with a third cell type, which multiplies the possibilities of analyzing more complex glomerular intercellular signaling events. In conclusion, the ease of assembly and versatility of use are the major advantages of this three-dimensional model of the glomerular filtration barrier over existing methods. The possibility to run a functional test that reliably measures albumin permeability makes the device a valid companion in several research applications ranging from drug screening to intercellular signaling studies.

Original languageEnglish
Pages (from-to)1-12
Number of pages12
JournalEuropean Journal of Pharmaceutical Sciences
Volume86
DOIs
Publication statusPublished - Apr 30 2016

Keywords

  • Albumin permeability
  • Drug testing
  • Endothelial cells
  • Glomerular filtration barrier
  • Intercellular signaling
  • Mesenchymal stem cells
  • Podocytes
  • Three-dimensional co-culture

ASJC Scopus subject areas

  • Pharmaceutical Science

Fingerprint Dive into the research topics of 'Three-dimensional podocyte-endothelial cell co-cultures: Assembly, validation, and application to drug testing and intercellular signaling studies'. Together they form a unique fingerprint.

  • Cite this