Conditioned Media From Glial Cells Promote a Neural-Like Connexin Expression in Human Adipose-Derived Mesenchymal Stem Cells.

Debora Lo Furno,Rosalba Parenti,Elisa Gili,Giuliana Mannino,Rosario Giuffrida,Agata Zappalà,Rosalia Pellitteri,Carlo Vancheri

doi:10.3389/fphys.2018.01742

Abstract

The expression of neuronal and glial connexins (Cxs) has been evaluated in adipose-derived mesenchymal stem cells (ASCs) whose neural differentiation was promoted by a conditioned medium (CM) obtained from cultures of olfactory ensheathing cells (OECs) or Schwann cells (SCs). By immunocytochemistry and flow cytometer analysis it was found that Cx43 was already considerably expressed in naïve ASCs and further increased after 24 h and 7 days from CM exposition. Cx32 and Cx36 were significantly improved in conditioned cultures compared to control ASCs, whereas a decreased expression was noticed in the absence of CM treatments. Cx47 was virtually absent in any conditions. Altogether, high basal levels and induced increases of Cx43 expression suggest a potential attitude of ASCs toward an astrocyte differentiation, whereas the lack of Cx47 would indicate a poor propensity of ASCs to become oligodendrocytes. CM-evoked Cx32 and Cx36 increases showed that a neuronal- or a SC-like differentiation can be promoted by using this strategy. Results further confirm that environmental cues can favor an ASC neural differentiation, either as neuronal or glial elements. Of note, the use of glial products present in CM rather than the addition of chemical agents to achieve such differentiation would resemble “more physiological” conditions of differentiation. As a conclusion, the overexpression of typical neural Cxs would indicate the potential capability of neural-like ASCs to interact with neighboring neural cells and microenvironment.

Highlights

Mesenchymal stem cells (MSCs) are adult stem cells featuring self-renewal and differentiation abilities
In accordance with previous studies (Calabrese et al, 2015), cells were immunopositive for typical MSC markers (CD 44, CD 73, CD 90, and CD 105) and immunonegative for typical hematopoietic stem cell markers (CD 14, CD 34, and CD 45)
As reported recently (Lo Furno et al, 2018), an adipose-derived MSCs (ASCs) neural-like phenotype by using olfactory ensheathing cells (OECs)-conditioned medium (CM) or Schwann cells (SCs)-CM was here confirmed by the increased immunopositivity for Nestin, PGP9.5, and GFAP (Figure 1)

Summary

Introduction

Mesenchymal stem cells (MSCs) are adult stem cells featuring self-renewal and differentiation abilities. They can typically give rise to mesodermal derivatives such as chondrocytes (Musumeci et al, 2011), osteocytes (Calabrese et al, 2015) or adipocytes (Lo Furno et al, 2013a) but, under appropriate conditions, they can transdifferentiate into epithelial or neural cells (Zuk et al, 2002). Safford et al (2002) were able to induce adipose-derived MSCs (ASCs) into a neural phenotype. Some transcription factors such as Pax, NeuroD1, Tbr, and Tbr, associated to different stages of neurogenesis, were found overexpressed in ASCs after neural induction (Cardozo et al, 2012)

Results

Discussion

Conclusion