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1 September 2000 ISOLATION AND SERUM-FREE CULTURE OF EPITHELIAL CELLS DERIVED FROM EPITHELIAL RESTS OF MALASSEZ IN HUMAN PERIODONTAL LIGAMENT
TAKENORI YAMANAKA, AKIHIKO SAKAMOTO, YOSHIHARU TANAKA, YAN ZHANG, YASUTAKA HAYASHIDO, SHIGEAKI TORATANI, YASUMASA AKAGAWA, TETSUJI OKAMOTO
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Abstract

To study biological character and function of epithelial rests of Malassez (ERM) in human periodontal ligament, we have developed a serum-free culture system of epithelial cells (ME) derived from ERM. The mitogenic effects of fibroblast growth factor (FGF)-1, FGF-2, and FGF-7/keratinocyte growth factor (KGF) on ME, human periodontal ligament-derived fibroblasts (PLF), human oral epithelial cells (OE), and human submandibular gland-derived epithelial cells (SGE) were investigated under a serum-free culture condition. FGF-1 and FGF-7/KGF stimulated the growth of both ME and SGE but FGF-2 had no effect. On the other hand, FGF-1, FGF-2, and FGF-7/KGF increased the OE proliferation. These results suggested that the divergent requirement of FGF ligands among these cells would be attributed to the different expression pattern of FGF receptor (FGFR) messenger ribonucleic acid (mRNA) isotypes. Therefore, we examined the expression of FGFR isotypes in these cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of ME- and SGE-derived mRNAs revealed the presence of mRNA encoding FGFR2-IIIb, a high affinity receptor for FGF-1 and FGF-7/KGF. However, no mRNAs for other FGFR isotypes were detected in both ME and SGE. On the contrary, OE expressed FGFR1-IIIc, FGFR3-IIIb, and FGFR4 mRNAs as well as FGFR2-IIIb. These results indicate that FGF binding sites on ME dominantly bind to FGF-1 and FGF-7/KGF, which transduce their signals via FGFR2-IIIb. Immunohistochemical analysis, PCR–Southern, ribonuclease protection assay (RPA), and Western blotting revealed that PLF expressed FGF-7/KGF mRNA and its peptide. These observations suggest that FGF-7/KGF might mediate epithelial–mesenchymal interactions between ME and PLF to maintain normal structure and function of periodontal ligament.

TAKENORI YAMANAKA, AKIHIKO SAKAMOTO, YOSHIHARU TANAKA, YAN ZHANG, YASUTAKA HAYASHIDO, SHIGEAKI TORATANI, YASUMASA AKAGAWA, and TETSUJI OKAMOTO "ISOLATION AND SERUM-FREE CULTURE OF EPITHELIAL CELLS DERIVED FROM EPITHELIAL RESTS OF MALASSEZ IN HUMAN PERIODONTAL LIGAMENT," In Vitro Cellular & Developmental Biology - Animal 36(8), 548-553, (1 September 2000). https://doi.org/10.1290/1071-2690(2000)036<0548:IASFCO>2.0.CO;2
Received: 1 March 2000; Accepted: 1 March 2000; Published: 1 September 2000
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KEYWORDS
epithelial rests of Malassez
fibroblast growth factor
fibroblast growth factor receptor
keratinocyte growth factor
periodontal ligament
serum-free culture
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