title: Co-culture with periodontal ligament stem cells enhanced osteoblastic differentiation of MC3T3-E1 cells and osteoclastic differentiation of RAW264.7 cells.
abstract: OBJECTIVE
  Periodontal ligament stem cells (PDLSCs) are characterized by having multipotential differentiation and immunoregulatory properties, which are the main mechanisms of PDLSCs-mediated periodontal regeneration. Periodontal or bone regeneration requires coordination of osteoblast and osteoclast, however, very little is known about the interactions between PDLSCs and osteoblast-like cells or osteoclast precursors. In this study, the indirect co-culture approach was introduced to preliminarily elucidate the effects of PDLSCs on differentiation of osteoblast-like cells and osteoclast precursors in vitro.
  
  
  METHODS
  Human PDLSCs were obtained from premolars extracted and their stemness was identified in terms of their colony-forming ability, proliferative capacity, cell surface epitopes and multi-lineage differentiation potentials. A noncontact co-culture system of PDLSCs and preosteoblastic cell line MC3T3-E1 or osteoclast precursor cell line RAW264.7 was established, and osteoblastic differentiation of MC3T3-E1 and osteoclastic differentiation of RAW264.7 were evaluated.
  
  
  RESULTS
  PDLSCs exhibited features of mesenchymal stem cells. Further investigation through indirect co-culture system showed that PDLSCs enhanced ALP activity, expressions of ALP, Runx2, BSP, OPN mRNA and BSP, OPN proteins and mineralization matrix deposition in MC3T3-E1. Meanwhile, they improved maturation of osteoclasts and expressions of TRAP, CSTK, TRAF6 mRNA and TRAP, TRAF6 proteins in RAW264.7.
  
  
  CONCLUSIONS
  PDLSCs stimulates osteoblastic differentiation of osteoblast precursors and osteoclastic differentiation of osteoclast precursors, at least partially, in a paracrine fasion.
authors.affiliation: School of Stomatology, Shandong UniversityJinan 250012, Shandong, China; Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Shandong UniversityJinan 250012, Shandong, China; Department of Stomatology, Qingdao Municipal HospitalQingdao 266011, Shandong, China.
authors.forename: Shulan
authors.initials: S
authors.lastname: Chen
authors.affiliation: School of Stomatology, Shandong UniversityJinan 250012, Shandong, China; Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Shandong UniversityJinan 250012, Shandong, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Ye
authors.affiliation: Department of Periodontology, Affiliated Hospital of Qingdao University Qingdao 266011, Shandong, China.
authors.forename: Xinbo
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Periodontology, Affiliated Hospital of Qingdao University Qingdao 266011, Shandong, China.
authors.forename: Quanchen
authors.initials: Q
authors.lastname: Xu
authors.affiliation: Department of Periodontology, Affiliated Hospital of Qingdao University Qingdao 266011, Shandong, China.
authors.forename: Keqing
authors.initials: K
authors.lastname: Pan
authors.affiliation: Department of Stomatology, Qingdao Municipal Hospital Qingdao 266011, Shandong, China.
authors.forename: Shulai
authors.initials: S
authors.lastname: Lu
authors.affiliation: School of Stomatology, Shandong UniversityJinan 250012, Shandong, China; Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Shandong UniversityJinan 250012, Shandong, China.
authors.forename: Pishan
authors.initials: P
authors.lastname: Yang
mesh_terms: D000293:Adolescent; D000328:Adult; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology*; D010010:Osteoclasts / Q000166:cytology*; D010513:Periodontal Ligament / Q000166:cytology*; D060888:Real-Time Polymerase Chain Reaction
keywords: Periodontal ligament stem cells; co-culture; osteoblastic differentiation; osteoclastic differentiation
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