Cysteamine treatment restores the in vitro ability to differentiate along the osteoblastic lineage of mesenchymal stromal cells isolated from bone marrow of a cystinotic patientReport as inadecuate




Cysteamine treatment restores the in vitro ability to differentiate along the osteoblastic lineage of mesenchymal stromal cells isolated from bone marrow of a cystinotic patient - Download this document for free, or read online. Document in PDF available to download.

Journal of Translational Medicine

, 13:143

First Online: 07 May 2015Received: 27 October 2014Accepted: 16 April 2015DOI: 10.1186-s12967-015-0494-0

Cite this article as: Conforti, A., Taranta, A., Biagini, S. et al. J Transl Med 2015 13: 143. doi:10.1186-s12967-015-0494-0

Abstract

BackgroundCystinosis is a rare autosomal recessive disease caused by mutations of the CTNS gene, which encodes for a lysosomal cystine-H symporter. In mice, inactivation of the CTNS gene causes intralysosomal cystine accumulation and progressive organ damage that can be reversed, at least in part, by infusion of mesenchymal stromal cells MSCs. Little is known on the mesenchymal compartment of cystinotic patients. The aim of the study was to test the phenotypical and functional properties of cystinotic MSCs Cys-MSCs isolated from bone marrow BM aspirate of a patient with nephropathic cystinosis.

MethodsMorphology, proliferative capacity measured as population doublings, immunophenotype by flow-cytometry and immunomodulatory properties as phytohemagglutinin-induced peripheral blood mononuclear cell proliferation were analyzed. The osteogenic differentiation potential of Cys-MSCs was evaluated by histological staining alkaline phosphatase activity, Alzarin Red and von Kossa staining spectrophotometry and Quantitative Reverse Transcriptase Polymerase Chain Reaction for osteigenic markers in the presence and in the absence of cysteamine. Cys-MSCs were compared with those isolated and expanded ex vivo from three healthy donors HD-MSCs.

ResultsDespite a slightly lower proliferative capacity, Cys-MSCs displayed a characteristic spindle-shaped morphology and similar immunephenotype as HD-MSCs. Cys-MSCs and HD-MSCs prevented proliferation of PHA-stimulated allogeneic peripheral blood mononuclear cells to the same extent. After in vitro induction into osteoblasts, Cys-MSCs showed reduced alkaline phosphatase ALP activity, calcium depositions and expression of ALP and collagen type 1. When Cys-MSCs were treated in vitro with increasing doses of cysteamine 50-100-200 μM-L during the differentiation assay, recovery of Cys-MSCs differentiation capacity into osteoblasts was observed. No difference in adipogenic differentiation was found between Cys-MSCs and HD-MSCs.

ConclusionsOur results indicate that, as compared to HD-MSCs, Cys-MSCs show reduced ability to differentiate into osteoblasts, which can be reverted after cysteamine treatment.

KeywordsMesenchymal stromal cells Cystinosis Osteogenic differentiation Cysteamine  Download fulltext PDF



Author: Antonella Conforti - Anna Taranta - Simone Biagini - Nadia Starc - Angela Pitisci - Francesco Bellomo - Valentina Cirillo -

Source: https://link.springer.com/







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