title: Expression of multiple stem cell markers in dental pulp cells cultured in serum-free media.
abstract: BACKGROUND
  Stem cell lines are usually grown in medium containing animal products. Fetal bovine serum (FBS) is an important additive for cell growth; however, the allergenic potential and the possibility of contamination when we use a medium containing serum would be a barrier to transplantation and consequently to the introduction of cell therapy methods into clinical applications.
  
  
  METHODS
  Dental mesenchymal cells were isolated and expanded in vitro and maintained in 4 different serum-free media (SFMs): SFM#1 (ITS-X, embryotrophic factor [ETF]); SFM#2 (ITS-X); SFM#3 (ETF); and SFM#4 (ETF, sodium pyruvate, ascorbic acid, fibroblast growth factor [FGF-a], acidic). Viability, proliferative, and immunocytochemical tests for the cells were performed by using 4 stem cell markers (CD44H, CK19, nestin, and P63) for ectoderm, mesoderm, and endoderm.
  
  
  RESULTS
  Viability tests showed a significant difference between the control and SFMs in both deciduous tooth pulp cells (DTPCs) and wisdom tooth pulp cells (WTPCs). However, all SFMs demonstrated 84%-90% viability, whereas the control showed 90%-93%. In both DTPCs and WTPCs, SFM#1 had the highest proliferation rate among the 4 SFMs. Immunocytochemistry stained positive stem cell markers most intensely in cells cultured with SFM#1. Furthermore, all stem cell markers for ectoderm, mesoderm, and endoderm were expressed in the cells cultured with SFM#1.
  
  
  CONCLUSIONS
  SFM#1 showed an acceptable survival rate, the highest proliferation rate, and the strongest expression of all the stem cell markers. SFM#1 proved to be a suitable medium for the culture of human dental pulp stem cells and to preserve pluripotency in differentiation.
authors.affiliation: Department of Oral Health, Nippon Dental University, School of Life Dentistry at Tokyo, Tokyo, Japan.
authors.forename: Thais Miyuki
authors.initials: TM
authors.lastname: Hirata
authors.forename: Nikolay
authors.initials: N
authors.lastname: Ishkitiev
authors.forename: Ken
authors.initials: K
authors.lastname: Yaegaki
authors.forename: Bogdan
authors.initials: B
authors.lastname: Calenic
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ishikawa
authors.forename: Taka
authors.initials: T
authors.lastname: Nakahara
authors.forename: Vanyo
authors.initials: V
authors.lastname: Mitev
authors.forename: Tomoko
authors.initials: T
authors.lastname: Tanaka
authors.forename: Markus
authors.initials: M
authors.lastname: Haapasalo
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis; D002417:Cattle; D002452:Cell Count; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media; D016895:Culture Media, Serum-Free; D003782:Dental Pulp / Q000166:cytology* / Q000187:drug effects; D004475:Ectoderm / Q000166:cytology; D004707:Endoderm / Q000166:cytology; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D006801:Humans; D018960:Hyaluronan Receptors / Q000032:analysis; D007328:Insulin / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D007381:Intermediate Filament Proteins / Q000032:analysis; D053539:Keratin-19 / Q000032:analysis; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000502:physiology*; D008648:Mesoderm / Q000166:cytology; D008964:Molar, Third / Q000166:cytology; D009419:Nerve Tissue Proteins / Q000032:analysis; D064231:Nestin; D019289:Pyruvic Acid / Q000494:pharmacology; D012643:Selenium / Q000494:pharmacology; D014094:Tooth, Deciduous / Q000166:cytology; D015534:Trans-Activators / Q000032:analysis; D014157:Transcription Factors; D014168:Transferrin / Q000494:pharmacology; D025521:Tumor Suppressor Proteins / Q000032:analysis
chemical_list: D015415:Biomarkers; C497483:CD44 protein, human; D003470:Culture Media; D016895:Culture Media, Serum-Free; D018960:Hyaluronan Receptors; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D007381:Intermediate Filament Proteins; D053539:Keratin-19; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; C115051:TP63 protein, human; D015534:Trans-Activators; D014157:Transcription Factors; D014168:Transferrin; D025521:Tumor Suppressor Proteins; D016220:Fibroblast Growth Factor 1; D019289:Pyruvic Acid; D012643:Selenium; D001205:Ascorbic Acid