Protocol to generate an in vitro model to study vascular calcification using human endothelial and smooth muscle cells

Elisa Persiani; Elisa Ceccherini; Ilaria Gisone; Antonella Cecchettini; Federico Vozzi

doi:10.1016/j.xpro.2023.102328

Protocol to generate an in vitro model to study vascular calcification using human endothelial and smooth muscle cells

STAR Protoc. 2023 Jun 9;4(2):102328. doi: 10.1016/j.xpro.2023.102328. Online ahead of print.

Authors

Elisa Persiani¹, Elisa Ceccherini², Ilaria Gisone³, Antonella Cecchettini⁴, Federico Vozzi⁵

Affiliations

¹ Institute of Clinical Physiology, National Research Council, Pisa, Italy. Electronic address: elisa.persiani@ifc.cnr.it.
² Institute of Clinical Physiology, National Research Council, Pisa, Italy. Electronic address: ceccherini@ifc.cnr.it.
³ Institute of Clinical Physiology, National Research Council, Pisa, Italy.
⁴ Institute of Clinical Physiology, National Research Council, Pisa, Italy; Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
⁵ Institute of Clinical Physiology, National Research Council, Pisa, Italy. Electronic address: vozzi@ifc.cnr.it.

Abstract

Vascular calcification is a systemic disease characterized by calcium salt deposition within vascular walls. Here, we present a protocol for establishing an advanced dynamic in vitro co-culture system using endothelial and smooth muscle cells to replicate vascular tissue complexity. We describe steps for cell culture and seeding in a double-flow bioreactor that recreates the action of blood in humans. We then detail the induction of calcification, setting up of the bioreactor, followed by cell viability assessment and calcium quantification.

Keywords: Biotechnology and bioengineering; Cell Biology; Tissue Engineering.