A reduction in CD90 (THY-1) expression results in increased differentiation of mesenchymal stromal cells.

Daniela A Moraes,Luciana C Marti,Ricardo B Azevedo,Paula Q Alvim,Jaqueline R Da Silva,Orlando A Toledo,Aline Pic-Taylor,Daniela M Oliveira,Tatiana T Sibov,Raphael S Bonadio,Lorena F Pavon

doi:10.1186/s13287-016-0359-3

Abstract

BackgroundMesenchymal stromal cells (MSCs) are multipotent progenitor cells used in several cell therapies. MSCs are characterized by the expression of CD73, CD90, and CD105 cell markers, and the absence of CD34, CD45, CD11a, CD19, and HLA-DR cell markers. CD90 is a glycoprotein present in the MSC membranes and also in adult cells and cancer stem cells. The role of CD90 in MSCs remains unknown. Here, we sought to analyse the role that CD90 plays in the characteristic properties of in vitro expanded human MSCs.MethodsWe investigated the function of CD90 with regard to morphology, proliferation rate, suppression of T-cell proliferation, and osteogenic/adipogenic differentiation of MSCs by reducing the expression of this marker using CD90-target small hairpin RNA lentiviral vectors.ResultsThe present study shows that a reduction in CD90 expression enhances the osteogenic and adipogenic differentiation of MSCs in vitro and, unexpectedly, causes a decrease in CD44 and CD166 expression.ConclusionOur study suggests that CD90 controls the differentiation of MSCs by acting as an obstacle in the pathway of differentiation commitment. This may be overcome in the presence of the correct differentiation stimuli, supporting the idea that CD90 level manipulation may lead to more efficient differentiation rates in vitro.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0359-3) contains supplementary material, which is available to authorized users.

Highlights

Mesenchymal stromal cells (MSCs) are multipotent progenitor cells used in several cell therapies
Transduction and establishment of small hairpin RNA (shRNA) (CD90 and control) expressing MSCs were performed using samples from all tissues, with similar levels of reduction in CD90 expression observed in all samples (Additional file 1: Table S1). quantitative real-time (qRT)- polymerase chain reaction (PCR) confirmed that shRNA CD90 used here effectively reduced transcript levels of CD90 (Fig. 1c)
Our results showed that the knockdown of CD90 leads to a decrease in CD44 and CD166 expression, which could indicate a shift in the stemness state of MSCs towards a state more susceptible to differentiation

Summary

Introduction

Mesenchymal stromal cells (MSCs) are multipotent progenitor cells used in several cell therapies. We sought to analyse the role that CD90 plays in the characteristic properties of in vitro expanded human MSCs. Mesenchymal stromal cells (MSCs) are multipotent progenitor cells identified by their plastic-adherence when maintained under standard culture conditions, selfrenewability, and differentiation into several mesodermal lineages [1,2,3]. MSCs are classically able to differentiate into osteoblasts, adipocytes, and chondroblasts in vitro [4] Since their initial description as colony-forming cell units present in the bone marrow [5], MSCs have been isolated from many tissue sources such as placenta [6], dental pulp [7], tendons [8], scalp tissue [9], adipose tissue [10], umbilical cord blood [11], umbilical cord perivascular. High CD90 expression has been related to the undifferentiated status of MSCs, since a decrease in CD90 level can be correlated with the temporal lineage commitment in vitro [25]

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Discussion

Conclusion