Journal Article

Use of dual-flow bioreactor to develop a simplified model of nervous-cardiovascular systems crosstalk: A preliminary assessment

Ghidoni, Alice, Marchesi, Nicoletta, Govoni, Stefano, Vanoli, Emilio, Parati, Gianfranco, Barbieri, Annalisa, Fahmideh, Foroogh, Pascale, Alessia, Calvillo, Laura

2020

Overview

Chronic conditions requiring long-term rehabilitation therapies, such as hypertension, stroke, or cancer, involve complex interactions between various systems/organs of the body and mutual influences, thus implicating a multiorgan approach. The dual-flow IVTech LiveBox2 bioreactor is a recently developed inter-connected dynamic cell culture model able to mimic organ crosstalk, since cells belonging to different organs can be connected and grown under flow conditions in a more physiological environment. This study aims to setup for the first time a 2-way connected culture of human neuroblastoma cells, SH-SY5Y, and Human Coronary Artery Smooth Muscle Cells, HCASMC through a dual-flow IVTech LiveBox2 bioreactor, in order to represent a simplified model of nervous-cardiovascular systems crosstalk, possibly relevant for the above-mentioned diseases. The system was tested by treating the cells with 10nM angiotensin II (AngII) inducing PKCβII/HuR/VEGF pathway activation, since AngII and PKCβII/HuR/VEGF pathway are relevant in cardiovascular and neuroscience research. Three different conditions were applied: 1- HCASMC and SH-SY5Y separately seeded in petri dishes (static condition); 2- the two cell lines separately seeded under flow (dynamic condition); 3- the two lines, seeded in dynamic conditions, connected, each maintaining its own medium, with a membrane as interface for biohumoral changes between the two mediums, and then treated. We detected that only in condition 3 there was a synergic AngII-dependent VEGF production in SH-SY5Y cells coupled to an AngII-dependent PKCβII/HuR/VEGF pathway activation in HCASMC, consistent with the observed physiological response in vivo . HCASMC response to AngII seems therefore to be generated by/derived from the reciprocal cell crosstalk under the dynamic inter-connection ensured by the dual flow LiveBox 2 bioreactor. This system can represent a useful tool for studying the crosstalk between organs, helpful for instance in rehabilitation research or when investigating chronic diseases; further, it offers the advantageous opportunity of cultivating each cell line in its own medium, thus mimicking, at least in part, distinct tissue milieu .

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Angiotensin / Angiotensin II / Biology and Life Sciences / Bioreactors / Biotechnology / Cell adhesion & migration / Cell Communication / Cell culture / Cell Line, Tumor / Cellular signal transduction / Chronic conditions / Chronic illnesses / Circulatory system / Coronary artery / Crosstalk / Health aspects / Humans / HuR protein / Hypertension / Laboratory animals / Medical research / Metabolism / Methods / Mimicry / Models, Cardiovascular / Models, Neurological / Myocytes, Smooth Muscle - cytology / Myocytes, Smooth Muscle - metabolism / Nervous system / Neuroblastoma / Neuroblastoma cells / Neuroblasts / Neurons - cytology / Neurons - metabolism / Organs / Pharmacology / Physical Sciences / Physiological aspects / Physiology / Rehabilitation / Research and analysis methods / Signal Transduction / Smooth muscle / Vascular endothelial growth factor