title: Differentiation of mesenchymal stem cells derived from pancreatic islets and bone marrow into islet-like cell phenotype.
abstract: BACKGROUND
  Regarding regenerative medicine for diabetes, accessible sources of Mesenchymal Stem Cells (MSCs) for induction of insular beta cell differentiation may be as important as mastering the differentiation process itself.
  
  
  RESULTS
  In the present work, stem cells from pancreatic islets (human islet-mesenchymal stem cells, HI-MSCs) and from human bone marrow (bone marrow mesenchymal stem cells, BM-MSCs) were cultured in custom-made serum-free medium, using suitable conditions in order to induce differentiation into Islet-like Cells (ILCs). HI-MSCs and BM-MSCs were positive for the MSC markers CD105, CD73, CD90, CD29. Following this induction, HI-MSC and BM-MSC formed evident islet-like structures in the culture flasks. To investigate functional modifications after induction to ILCs, ultrastructural analysis and immunofluorescence were performed. PDX1 (pancreatic duodenal homeobox gene-1), insulin, C peptide and Glut-2 were detected in HI-ILCs whereas BM-ILCs only expressed Glut-2 and insulin. Insulin was also detected in the culture medium following glucose stimulation, confirming an initial differentiation that resulted in glucose-sensitive endocrine secretion. In order to identify proteins that were modified following differentiation from basal MSC (HI-MSCs and BM-MSCs) to their HI-ILCs and BM-ILCs counterparts, proteomic analysis was performed. Three new proteins (APOA1, ATL2 and SODM) were present in both ILC types, while other detected proteins were verified to be unique to the single individual differentiated cells lines. Hierarchical analysis underscored the limited similarities between HI-MSCs and BM-MSCs after induction of differentiation, and the persistence of relevant differences related to cells of different origin.
  
  
  CONCLUSIONS
  Proteomic analysis highlighted differences in the MSCs according to site of origin, reflecting spontaneous differentiation and commitment. A more detailed understanding of protein assets may provide insights required to master the differentiation process of HI-MSCs to functional beta cells based only upon culture conditioning. These findings may open new strategies for the clinical use of BM-MSCs in diabetes.
authors.affiliation: Molecular Biotechnology Centre, University of Turin, Turin, Italy. cristina.zanini@unito.it
authors.forename: Cristina
authors.initials: C
authors.lastname: Zanini
authors.forename: Stefania
authors.initials: S
authors.lastname: Bruno
authors.forename: Giorgia
authors.initials: G
authors.lastname: Mandili
authors.forename: Denisa
authors.initials: D
authors.lastname: Baci
authors.forename: Francesco
authors.initials: F
authors.lastname: Cerutti
authors.forename: Giovanna
authors.initials: G
authors.lastname: Cenacchi
authors.forename: Leo
authors.initials: L
authors.lastname: Izzi
authors.forename: Giovanni
authors.initials: G
authors.lastname: Camussi
authors.forename: Marco
authors.initials: M
authors.lastname: Forni
mesh_terms: D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D002469:Cell Separation; D048430:Cell Shape / Q000187:drug effects; D016000:Cluster Analysis; D003470:Culture Media / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure*; D010641:Phenotype; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D003470:Culture Media; D007328:Insulin