Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Oct 26, 2020; 12(10): 1152-1170
Published online Oct 26, 2020. doi: 10.4252/wjsc.v12.i10.1152
Pericyte-like differentiation of human adipose-derived mesenchymal stem cells: An in vitro study
Giuliana Mannino, Florinda Gennuso, Giovanni Giurdanella, Federica Conti, Filippo Drago, Salvatore Salomone, Debora Lo Furno, Claudio Bucolo, Rosario Giuffrida
Giuliana Mannino, Debora Lo Furno, Rosario Giuffrida, Physiology Section, Department of Biomedical and Biotechnological Sciences, University of Catania, Catania 95123, Italy
Florinda Gennuso, Federica Conti, Filippo Drago, Salvatore Salomone, Claudio Bucolo, Pharmacology Section, Department of Biomedical and Biotechnological Sciences, School of Medicine, University of Catania, Catania 95123, Italy
Giovanni Giurdanella, Biochemistry Section, Department of Biomedical and Biotechnological Sciences, School of Medicine, University of Catania, Catania 95123, Italy
Author contributions: Mannino G and Gennuso F made substantial contributions to conception and design, participated in the acquisition, analysis and interpretation of data; Giurdanella G and Conti F participated in data acquisition; Salomone S and Drago F were involved in data analysis and interpretation, and revising the manuscript critically for important intellectual content; Lo Furno D, Bucolo C and Giuffrida R conceived the project and developed the experimental design, analyzed data and wrote the manuscript. All authors reviewed the manuscript and approved the version to be published.
Supported by “Piano Triennale per la Ricerca 2016-2018 – Linea Intervento 2”, University of Catania, Italy, No. 20722142118.
Institutional review board statement: This study was reviewed and approved by the local ethics committee (Comitato etico Catania1; Authorization n. 155/2018/PO).
Conflict-of-interest statement: The authors declare no conflict of interest.
Data sharing statement: No additional data are available.
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 NonCommercial (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: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Debora Lo Furno, PhD, Assistant Professor, Physiology Section, Department of Biomedical and Biotechnological Sciences, University of Catania, No. 95125 Catania, Catania 95123, Italy. lofurno@unict.it
Received: May 15, 2020
Peer-review started: May 15, 2020
First decision: June 7, 2020
Revised: June 18, 2020
Accepted: August 25, 2020
Article in press: August 25, 2020
Published online: October 26, 2020
ARTICLE HIGHLIGHTS
Research background

The loss of pericytes, which occurs in diabetic retinopathy, results in a breakdown of the blood-retina barrier (BRB) and infiltration of inflammatory cells. Lost retinal pericytes might be replaced by adipose-derived mesenchymal stem cells (ASCs) after differentiating into a pericyte-like phenotype.

Research motivation

The use of pericyte-like differentiated ASCs may represent a valuable therapeutic strategy for restoring BRB damage.

Research objectives

The purpose of this study was to develop in vitro strategies to obtain pericyte-like differentiation of human ASCs.

Research methods

Different ASC culture conditions were tested and compared to human retinal pericytes (hRPCs). The expression of α-smooth muscle actin (α-SMA) and neural/glial antigen 2 (NG2) was assessed by immunocytochemical staining and western blotting. In co-cultures of human retinal endothelial cells (hRECs) with hRPCs or different groups of hASCs, the endothelial expression of typical junctional proteins such as vascular endothelial-Cadherin, zonula occludens-1 and Occludin were evaluated. In an in vitro model of BRB, the trans-endothelial electrical resistance was measured. Three-dimensional co-cultures in Matrigel of hRECs and hRPCs or pericyte-like hASCs were designed to assess their reciprocal localization.

Research results

Immunocytochemical results and western blot analysis for α-SMA and NG2 indicated that the closest pericyte-like phenotype was observed when hASCs were cultured in pericyte medium (PM-hASCs). α-SMA immunoreactivity was strongly increased only when TGF was added to the culture medium. NG2 expression, almost undetectable in most conditions, was substantially increased in PM-hASCs.

In co-culture experiments, immunoreactivity of vascular endothelia-Cadherin, zonula occludens-1 and Occludin was considerably increased in hRECs when hRPCs or PM-hASCs were also present. Supporting results were found by trans-endothelial electrical resistance measurements, showing the highest values in analogous conditions. The pericyte-like phenotype of PM-hASCs was also confirmed in three-dimensional co-cultures in Matrigel, where PM-hASCs and hRPCs similarly localized around the tubular formations made by hRECs.

Research conclusions

PM-hASCs seem able to strengthen the intercellular junctions between hRECs, likely reinforcing the BRB; thus, hASC-based therapeutic approaches may be usefully developed to restore the integrity of retinal microcirculation.

Research perspectives

Future in vivo experiments will be designed to test pericyte-like ASC engraftment following intraocular administration. Possible beneficial effects will be evaluated in animal models of diabetic retinopathy. Eventually, when safe therapeutic protocols are developed, ASC-based therapeutic approaches may be successfully used in diabetic patients to restore disrupted retinal microcirculation.