title: Optimal medium formulation for the long-term expansion and maintenance of human periodontal ligament stem cells.
abstract: BACKGROUND
  Human periodontal ligament stem cells (hPDLSCs) are promising mesenchymal stem cells that are readily accessible. However, there is as yet no consensus as to the optimal culture medium for hPDLSCs. Thus, the purpose of the present study is to determine the optimal culture medium for long-term expansion of hPDLSCs.
  
  
  METHODS
  hPDLSCs were isolated from healthy third molars, and the most widely used medium formulations in previous studies were used: 1) an α minimum essential medium-based medium formulation (MBM); and 2) a Dulbecco's minimum essential medium-based medium formulation. Passage 5 (P5) and P8 were evaluated with the two media for cell proliferation, differentiation, and immunophenotype.
  
  
  RESULTS
  hPDLSCs that were primarily cultured in MBM were far more proliferated than those grown in DBM. In general, application of the MBM for longer periods produced greater cell growth and osteogenic differentiation. Furthermore, MBM-precultured hPDLSCs exhibited a greater degree of cell proliferation and a greater production of mineralized tissue and alkaline phosphatase (ALP) activity in vitro, although the levels of both were dependent on the culture medium used. With respect to long-term expansion, the P5 hPDLSCs grew and produced the largest amount of mineralized nodules faster than the P8 hPDLSCs, but both passages exhibited a similar phenotype for stemness and ALP activity.
  
  
  CONCLUSIONS
  The present study indicates that the inherent capacity of hPDLSCs could be maintained until a later passage, P8 in MBM, and MBM appears to be an optimal choice for manipulating the finest and most stable hPDLSCs.
authors.affiliation: Division of Periodontology, Department of Dentistry, School of Medicine, Inha University, Incheon, South Korea.
authors.forename: Im-Hee
authors.initials: IH
authors.lastname: Jung
authors.forename: Bo-Sun
authors.initials: BS
authors.lastname: Kwon
authors.forename: Se-Hee
authors.initials: SH
authors.lastname: Kim
authors.forename: Hye-Eun
authors.initials: HE
authors.lastname: Shim
authors.forename: Choong-Man
authors.initials: CM
authors.lastname: Jun
authors.forename: Jeong-Ho
authors.initials: JH
authors.lastname: Yun
mesh_terms: D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis; D015703:Antigens, CD / Q000032:analysis; D018941:Antigens, CD19 / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D051929:CD146 Antigen / Q000032:analysis; D002113:Calcification, Physiologic / Q000502:physiology; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D048430:Cell Shape / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002626:Chemistry, Pharmaceutical; D003470:Culture Media / Q000737:chemistry*; D000071063:Endoglin; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D018960:Hyaluronan Receptors / Q000032:analysis; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D010012:Osteogenesis / Q000502:physiology; D010513:Periodontal Ligament / Q000166:cytology*; D010641:Phenotype; D011956:Receptors, Cell Surface / Q000032:analysis; D013997:Time Factors
chemical_list: D015703:Antigens, CD; D018941:Antigens, CD19; D000954:Antigens, Surface; D051929:CD146 Antigen; C497483:CD44 protein, human; D003470:Culture Media; C064206:ENG protein, human; D000071063:Endoglin; D018960:Hyaluronan Receptors; C497902:MCAM protein, human; D011956:Receptors, Cell Surface; C525591:STRO-1 antigen, human; D000469:Alkaline Phosphatase