Effects of lactoferrin on the differentiation of pluripotent mesenchymal cells

Authors

  • Motohiko Yagi,

    1. Nihon University Graduate School of Dentistry, Tokyo, Japan
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  • Naoto Suzuki,

    Corresponding author
    1. Department of Biochemistry, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
    2. Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
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  • Tadahiro Takayama,

    1. Department of Periodontology, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
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  • Masatoshi Arisue,

    1. Department of Biochemistry, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
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  • Takuya Kodama,

    1. Department of Biochemistry, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
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  • Yasushi Yoda,

    1. Department of Biochemistry, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
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  • Kichibee Otsuka,

    1. Department of Biochemistry, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
    2. Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
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  • Koichi Ito

    1. Department of Periodontology, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
    2. Division of Clinical Research, Dental Research Center, Nihon University School of Dentistry, Tokyo, Japan
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Department of Biochemistry, Nihon University School of Dentistry, 1-8-13, Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan. Tel.: +81 3 3219 8123; fax: +81 3 3219 8334. E-mail addresses: suzuki-n@dent.nihon-u.ac.jp

Abstract

Lactoferrin accelerates bone formation, but the precise cellular mechanism behind this is still unclear. We examined the effect of lactoferrin on the differentiation of pluripotent mesenchymal cells using a typical pluripotent mesenchymal cell line, C2C12. Cells were cultured in low-mitogen differentiation medium to induce cell differentiation, with or without the addition of lactoferrin. The cell lineage was determined by alkaline phosphatase (ALPase) activity, mRNA expression of cellular phenotype-specific markers using real-time polymerase chain reaction (PCR), and protein synthesis using Western blotting. The expression of low-density lipoprotein lipase receptor-related proteins (LRPs) 1 and 2, both lactoferrin receptors, was determined by reverse transcription-PCR. ALPase activity increased after the addition of lactoferrin. The mRNA expression of Runx2, osteocalcin, and Sox9 increased markedly as a result of lactoferrin treatment, whereas the expression of MyoD, desmin, and PPARγ decreased significantly. Western blots showed that lactoferrin stimulation increased Runx2 and Sox9 proteins, whereas it decreased MyoD and PPARγ synthesis. C2C12 cells expressed the LRP1 lactoferrin receptor. These results indicate that lactoferrin treatment converts the differentiation pathway of C2C12 cells into the osteoblastic and chondroblastic lineage.

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