Effects of basic fibroblast growth factor on the development of the stem cell properties of human dental pulp cells.
We isolated adherent fibroblastic cells after collagenase and dispase treatment of human dental pulp. When human dental pulp cells (hDPCs) were cultured in the presence of basic fibroblast growth factor (bFGF), the ratio of hDPCs in the S-phase was significantly higher in comparison with incubation without bFGF. The ratio of hDPCs expressing STRO-1 as a marker of stem cell populations increased approximately eightfold in the presence of bFGF as opposed to that in the absence of bFGF. We demonstrated the characterization and distinctiveness of the hDPCs and showed that, when cultured with the medium containing serum and bFGF, they were highly proliferative and capable of differentiating in vitro into osteoblasts, chondrocytes, and adipocytes. Furthermore, the in vitro differentiation was confirmed at both the protein and gene expression levels. Transplantation of hDPCs -- expanded ex vivo in the presence of bFGF into immunocompromised mice -- revealed the formation of bone, cartilage, and adipose tissue. The donor hDPC-derived cells were labeled in the bone tissues located near the PLGA in the subcutaneous tissues of recipient mice using a human-specific Alu probe. When cultured with a serum-free medium containing bFGF, the hDPCs strongly expressed STRO-1 immunoreactive products and sustained self-renewal, and thus were almost identical in differentiation potential and proliferation activity to hDPCs cultured with the medium containing serum and bFGF. The present results suggest that the hDPCs cultured in the presence of bFGF irrespective of the presence or absence of the bovine serum are rich in mesenchymal stem cells or progenitor cells and useful for cell-based therapies to treat dental diseases.
Morito
Akiyuki
A
Department of Anatomy II, Tsurumi University School of Dental Medicine, Yokohama, Japan.
Kida
Yujiro
Y
Suzuki
Kazunori
K
Inoue
Koji
K
Kuroda
Noriyuki
N
Gomi
Kazuhiro
K
Arai
Takashi
T
Sato
Tetsuji
T
D050156:Adipogenesis / Q000187:drug effects; D000293:Adolescent; D000328:Adult; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003782:Dental Pulp / Q000166:cytology* / Q000201:enzymology / Q000637:transplantation / Q000648:ultrastructure; D050556:Fatty Acid-Binding Proteins / Q000235:genetics / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D017403:In Situ Hybridization; D047569:Laser Scanning Cytometry; D051379:Mice; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects*; D055815:Young Adult
D053674:Aggrecans; D000954:Antigens, Surface; D050659:Core Binding Factor Alpha 1 Subunit; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; C493410:FABP4 protein, human; D050556:Fatty Acid-Binding Proteins; C493709:RUNX2 protein, human; C525591:STRO-1 antigen, human; D016222:Fibroblast Growth Factor 2; D015675:Osteocalcin; D000469:Alkaline Phosphatase