Tissue homeostasis of intervertebral disc

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K10420
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Keio University
• Principal Investigator: Fujita Nobuyuki 慶應義塾大学, 医学部(信濃町), 講師 (30348685)
• Co-Investigator(Kenkyū-buntansha): 松本 守雄 慶應義塾大学, 医学部(信濃町), 教授 (40209656)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 椎間板 / 髄核 / 繊維輪 / 酸化ストレス / 細胞株

Outline of Final Research Achievements

We show that a chordoma-derived cell line U-CH1-N cells highly express the nucleus pulposus (NP) marker genes, including T, KRT19, and CD24. These observations were further confirmed by immunocytochemistry and flow cytometry. Reporter analyses showed that transcriptional activity of T was enhanced in U-CH1-N cells. Chondrogenic capacity of U-CH1-N cells was verified by evaluating the expression of ECM genes and Alcian blue staining. Of note, we found that proliferation and synthesis of chondrogenic ECM proteins were largely dependent on T in U-CH1-N cells. In accordance, knockdown of the T transcripts suppressed the expression of PCNA, a gene essential for DNA replication, and SOX5 and SOX6, the master regulators of chondrogenesis. On the other hand, the CD24-silenced cells showed no reduction in the mRNA expression level of the chondrogenic ECM genes. These results suggest that U-CH1-N shares important biological properties with notochordal NP cells.

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