Growth and differentiation of periodontal ligament-derived cells in serum-free defined culture

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Publisher: Springer
Document Type: Article
Length: 374 words

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Byline: Tetsuji Okamoto (1), Nobuhiro Yatsuzuka (2), Yoshiharu Tanaka (1), Mikio Kan (3), Takenori Yamanaka (2), Akihiko Sakamoto (3), Takashi Takata (4), Yasumasa Akagawa (2), Gordon H. Sato (5), J. Denry Sato (5), Kazuaki Takada (1) Keywords: periodontal ligament cells; serum-free culture; FGF-1; FGF-2; FGFR-1; alkaline phosphatase Abstract: We have developed a serum-free medium for the growth and differentiation of periodontal ligament-derived cells (PLC). In addition, the expression of both fibroblast growth factor (FGF) and FGF receptor (FGFR) in the PLC was investigated by immunohistochemical examination, heparin affinity chromatography (HAC), and reverse transcription-polymerase chain reaction (RT-PCR) analysis. Optimal growth of the cells was achieved in Iscove's modified Dulbecco's medium supplemented with insulin, transferrin, 2-mercaptoethanol, 2-ethanolamine, sodium selenite, and oleic acid in type-I collagen-coated dishes. Both FGF-1 and FGF-2 stimulated cell growth and inhibited differentiation as measured by inhibition of alkaline phosphatase activity of the cells. An immunohistochemical analysis of FGF-1 and FGF-2 revealed that immunoreactive FGF-1 and FGF-2 were detected predominantly in the cytoplasm of growing cells. In addition, perinuclear FGF-1 staining and nuclear FGF-2 staining were observed in the same growing cells. In contrast, a faint diffuse staining of FGF-1 and FGF-2 was detected in cytoplasm of the confluent differentiated cells. The 2.15 M NaCl eluate from HAC of the cell extracts exhibited growth-promoting activities for the PLC, and it also stimulated the growth of human umbilical vein-derived endothelial cells and inhibited binding of [[.sup.125]I]-FGF to its receptors, indicating the cells produced FGFs or FGF-like growth factors. RT-PCR analysis revealed that the cells expressed FGFR-1 mRNA but not mRNAs for FGFR-2, FGFR-3 and FGFR-4 mRNA. These results suggest that the FGF-FGFR-1 system plays an important role in the growth and differentiation of periodontal ligament-derived cells. Author Affiliation: (1) Department of Oral & Maxillofacial Surgery I, School of Dentistry, Hiroshima University, 1-2-3, Kasumi, Minami-ku, 734, Hiroshima, Japan (2) Department of Prosthetic Dentistry I, School of Dentistry, Hiroshima University, 734, Hiroshima, Japan (4) Department of Oral Pathology, School of Dentistry, Hiroshima University, 734, Hiroshima, Japan (3) Center for Cancer Biology and Nutrition, Institute of Bioscience and Technology, Texas A&M University, 77030-3303, Houston, Texas (5) W. Alton Jones Cell Science Center, Lake Placid, New York Article History: Registration Date: 17/05/1997 Received Date: 24/02/1996 Accepted Date: 05/03/1996

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Gale Document Number: GALE|A206211748