Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Sep 7, 2019; 25(33): 4892-4903
Published online Sep 7, 2019. doi: 10.3748/wjg.v25.i33.4892
Ex vivo effect of vascular wall stromal cells secretome on enteric ganglia
Giovanni Dothel, Chiara Bernardini, Augusta Zannoni, Maria Rosaria Spirito, Roberta Salaroli, Maria Laura Bacci, Monica Forni, Fabrizio De Ponti
Giovanni Dothel, Maria Rosaria Spirito, Fabrizio De Ponti, Department of Medical and Surgical Sciences, University of Bologna, Bologna 40126, Italy
Chiara Bernardini, Augusta Zannoni, Roberta Salaroli, Maria Laura Bacci, Monica Forni, Department of Veterinary Medical Sciences, University of Bologna, Bologna 40064, Italy
Author contributions: Dothel G, Zannoni A, Bernardini C and Forni M conceived and designed the study; Dothel G, Salaroli R and Spirito MR performed the experiments; Dothel G, Bernardini C and Forni M wrote the paper; Dothel G performed the data analysis; Bacci ML carried out the procedures on pigs; De Ponti F and Forni M coordinated the research; all authors reviewed and participated to the paper drafting.
Supported by Fondazione del Monte di Bologna e Ravenna (ID ROL: FdM/3208).
Institutional review board statement: This study was approved by the institutional review board of National Institute of Health.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Bologna (approved experimental protocol for the use of guinea pigs: protocol number - 18/79/14; approved experimental protocol for the use of pigs: protocol number - 43-IX/9 all.37; 15/04/2013).
Conflict-of-interest statement: None of the authors have any potential conflicts of interest associated with this research.
ARRIVE guidelines statement: The present study was conceived, designed and performed following the ARRIVE guidelines. All the authors have read the ARRIVE guidelines and reviewed the manuscript accordingly.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Corresponding author: Monica Forni, PhD, Associate Professor, Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra 50, Ozzano dell'Emilia, Bologna 40064, Italy.
Telephone: +39-51-2097913 Fax: +39-51-2097899
Received: April 11, 2019
Peer-review started: April 11, 2019
First decision: May 17, 2019
Revised: May 31, 2019
Accepted: June 8, 2019
Article in press: June 8, 2019
Published online: September 7, 2019
Research background

There is growing interest on mesenchymal stromal cells (MSC) as a novel therapeutic strategy to treat auto-immune and inflammatory diseases. However, identifying optimal MSC sources and limited reliability of current experimental models still represent a challenge in this field. Pigs represent more closely human physiology and an accessible resource for ex vivo procedures. Recently, our group isolated a population of pericytes from porcine aortic wall with an MSC profile, currently cited as porcine vascular wall-MSC (pVW-MSC).

Research motivation

Inflammatory bowel diseases (IBDs), comprising the two major forms ulcerative colitis and Crohn’s disease, are characterized by an aberrant immune response leading to severe damage of the intestinal wall and functioning. Current trials are evaluating the application of cell-based therapies for the treatment of IBDs. The present study describes the effect of pVW-MSC-conditioned medium (CM) on enteric ganglia in two ex vivo models of IBDs in order to investigate a potential development of MSC-based treatment of IBDs.

Research objective

To evaluate the effect of pVW-MSC secretome on survival and differentiation of enteric ganglionic cells isolated by guinea pigs (GPEG) and pigs (PEG) and exposed to lipopolysaccharide (LPS).

Research methods

The expression of standard MSC markers in pVW-MSC were assessed by flow cytometry. Increasing concentration of LPS were tested in both GPEG and PEG cultures. CM derived by pVW-MSC cultures were added alone or in combination with 1µg of LPS in GPEG and PEG cultures. Ganglionic cells were double-stained with antibodies directed to the pan-neuronal marker, HuD and the glial fibrillary acidic protein, GFAP. Cell count and morphometric analysis were performed to determine changes of neuronal and glial population.

Research results

Guinea-pig neurons and glial cells decreased and increased respectively in response to high concentrations of LPS. These changes were not observed in pig primary cultures. pVW-MSC secretome increased the number and differentiation of glial cells compared to neurons with a more pronounced effect in PEG and in combination with LPS.

Research conclusions

These data showed a higher resilience of pig enteric ganglia to the main bacterial product LPS compared to guinea pig and a higher responsiveness of glial cells to pVW-MSC secreted mediators.

Research perspectives

Neuro-immune changes induced by pVW-MSC represent an essential aspect in the development of cell-based therapies. Further studies are warranted to investigate inter-species differences of pVW-MSC secretome.