title: Low-serum culture with novel medium promotes maxillary/mandibular bone marrow stromal cell proliferation and osteogenic differentiation ability.
abstract: OBJECTIVE
  The purpose of this study was to evaluate the effect of low-serum STK2 medium on the isolation and osteogenic differentiation of human maxillary/mandibular bone marrow stromal cells (MBMSCs).
  
  
  METHODS
  Human MBMSCs were obtained from patients undergoing dental implant treatment. These cells were cultured in serum-free medium or STK2 medium containing 1  % fetal bovine serum (low-serum) or α-MEM containing 10  % fetal bovine serum (control). Proliferation on the culture surface, cell surface antigen expression, and mRNA levels of neural crest and osteogenic markers were examined. Alkaline phosphatase assay and Alizarin red staining were used to assess osteogenic differentiation potential. Immunoblotting analysis was performed to detect ERK phosphorylation.
  
  
  RESULTS
  Low-serum and control MBMSCs were positive for CD73, CD90, and CD105 and negative for CD14, CD34, CD45, CD271, and HLA-DR. CD140a was absent in low-serum cells but present in control cells. Low-serum MBMSCs proliferated more than control MBMSCs. After induction of osteogenic differentiation, alkaline phosphatase activity and osteocalcin mRNA levels were higher in low-serum MBMSCs than in control cells, and Alizarin red staining was stronger in low-serum MBMSCs than in control cells. Low-serum culture promoted ERK phosphorylation.
  
  
  CONCLUSIONS
  MBMSCs precultured in low-serum medium exhibited a greater cumulative cell number and a higher osteogenic differentiation capacity than those cultured in control medium.
  
  
  CONCLUSIONS
  These findings indicate that low-serum STK2 culture might be useful to promote MBMSC proliferation and osteogenic differentiation. This method requires less autologous blood collection for cell expansion than conventional methods, thus reducing the burden on patients.
authors.affiliation: Department of Oral and Maxillofacial Prosthodontics, Kagoshima University Graduate School of Biomedical Sciences, Kagoshima, Japan.
authors.forename: Fumio
authors.initials: F
authors.lastname: Suehiro
authors.affiliation: Department of Oral and Maxillofacial Prosthodontics, Kagoshima University Graduate School of Biomedical Sciences, Kagoshima, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Ishii
authors.affiliation: Department of Regenerative Oral Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Izumi
authors.initials: I
authors.lastname: Asahina
authors.affiliation: Department of Prosthetic Dentistry, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Murata
authors.affiliation: Department of Oral and Maxillofacial Prosthodontics, Kagoshima University Graduate School of Biomedical Sciences, Kagoshima, Japan. mnishi@dent.kagoshima-u.ac.jp.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nishimura
mesh_terms: D000954:Antigens, Surface / Q000032:analysis; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D003864:Depression, Chemical; D006801:Humans; D015151:Immunoblotting; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010012:Osteogenesis / Q000187:drug effects*; D010766:Phosphorylation; D012333:RNA, Messenger / Q000032:analysis; D013268:Stimulation, Chemical
chemical_list: D000954:Antigens, Surface; D016895:Culture Media, Serum-Free; D012333:RNA, Messenger
keywords: Bone tissue engineering; Low-serum medium; Maxillary/mandibular bone marrow stromal cells; Osteogenic differentiation