title: Proteomic analysis of stromal cells derived from the dental pulp of human exfoliated deciduous teeth.
abstract: Human dental pulp derived from exfoliated deciduous teeth has been described as a promising alternative source of multipotent stem cells. While these cells share certain similarities with mesenchymal stem-like cells (MSC) isolated from other tissues, basically they are still poorly characterized. In this study, for the first time, a proteomic map of abundantly expressed proteins in stromal cells derived from the dental pulp of human exfoliated deciduous teeth (SHED) was established. We also analyzed proteomic signatures of 2 clonal strains derived from SHEDs by single-cell cloning. The SHEDs were established from enzyme-disaggregated deciduous dental pulp from 6-year-old children. They had typical fibroblastoid morphology and high colony-forming efficiency index (16.4%). Cloning was performed at the second passage using limiting dilution in a 96-well plate (0.3 cell/well). Differentiation assessment revealed strong osteogenic but no adipogenic potential of the SHEDs in either clonal strain. The cells expressed characteristic antigens of MSC-like cells, including CD73, CD90, CD105, CD146, and did not express hematopoietic markers CD14, CD34, and CD45, as assessed with FACS analysis. For proteomic studies, cytosolic and nuclear proteins were analyzed with 2-dimensional gel electrophoresis (2-DE) and identified using matrix-assisted laser desorption/ionization (MALDI)-time of fl ight (TOF)-mass spectrometry (MS). All proteins were identified with high level of confidence (the lowest sequence coverage was 27%). Identification of highly expressed proteins in SHEDs revealed proteomic profiles very similar to that of MSC-like cells derived from other tissues. We also found a high degree of similarity between proteomic signatures of primary SHEDs and clonal cell strains. Thus, our data confirm a close resemblance between SHEDs and MSC-like cells from other tissues and may serve as starting point for creating-comprehensive proteomic maps.
authors.affiliation: Department of Experimental Medicine, Institute of Experimental and Clinical Medicine , Vilnius, Lithuania. augustas.pivoriunas@ekmi.vu.lt
authors.forename: Augustas
authors.initials: A
authors.lastname: Pivoriuūnas
authors.forename: Andrejus
authors.initials: A
authors.lastname: Surovas
authors.forename: Veronika
authors.initials: V
authors.lastname: Borutinskaite
authors.forename: Dalius
authors.initials: D
authors.lastname: Matuzeviccius
authors.forename: Graina
authors.initials: G
authors.lastname: Treigyte
authors.forename: Juūrate
authors.initials: J
authors.lastname: Savickiene
authors.forename: Virginijus
authors.initials: V
authors.lastname: Tunaitis
authors.forename: Ruūta
authors.initials: R
authors.lastname: Aldonyte
authors.forename: Akvile
authors.initials: A
authors.lastname: Jarmalavicciuūte
authors.forename: Kristina
authors.initials: K
authors.lastname: Suriakaite
authors.forename: Evaldas
authors.initials: E
authors.lastname: Liutkeviccius
authors.forename: Algirdas
authors.initials: A
authors.lastname: Venalis
authors.forename: Dalius
authors.initials: D
authors.lastname: Navakauskas
authors.forename: Ruūta
authors.initials: R
authors.lastname: Navakauskiene
authors.forename: Karl-Eric
authors.initials: KE
authors.lastname: Magnusson
mesh_terms: D050156:Adipogenesis / Q000502:physiology; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D002648:Child; D003782:Dental Pulp / Q000166:cytology*; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008969:Molecular Sequence Data; D039902:Multipotent Stem Cells / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D010012:Osteogenesis / Q000502:physiology; D020543:Proteome / Q000032:analysis*; D017154:Stromal Cells / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D014094:Tooth, Deciduous / Q000166:cytology*
chemical_list: D015415:Biomarkers; D020543:Proteome