Back to Search
Start Over
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
- Source :
- Journal of Translational Medicine
- Publication Year :
- 2015
- Publisher :
- BioMed Central Ltd., 2015.
-
Abstract
- Background: Cystinosis 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. Methods: Morphology, 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). Results: Despite 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. Conclusions: Our results indicate that, as compared to HD-MSCs, Cys-MSCs show reduced ability to differentiate into osteoblasts, which can be reverted after cysteamine treatment.
- Subjects :
- Pathology
medicine.medical_specialty
Adolescent
Cysteamine
Cystinosis
Cystine
Cell Culture Techniques
Mesenchymal stromal cells
Biology
General Biochemistry, Genetics and Molecular Biology
Immunophenotyping
chemistry.chemical_compound
Young Adult
Nephropathic Cystinosis
Bone Marrow
Osteogenic differentiation
medicine
Humans
Cell Lineage
Child
Cell Proliferation
Medicine(all)
Osteoblasts
Biochemistry, Genetics and Molecular Biology(all)
Research
Mesenchymal stem cell
Cell Differentiation
Mesenchymal Stem Cells
General Medicine
medicine.disease
3. Good health
medicine.anatomical_structure
chemistry
Settore MED/38 - PEDIATRIA GENERALE E SPECIALISTICA
Symporter
Cancer research
Leukocytes, Mononuclear
Bone marrow
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Journal of Translational Medicine
- Accession number :
- edsair.doi.dedup.....d70cb892990403c147f809e869edb9cd