我々はU-CH1-N細胞株を用いて, 髄核細胞との類似性を検証した結果, U-CH1-Nは髄核の分子マーカーであるBrachyury (T), CD24, Cytokeratin19の発現が高く, 更には軟骨基質であるAggrecanやType II collagenの発現も高く, 長期に培養したところ, アルシアンブルー陽性であり, 脊索由来の髄核細胞と分子生物学的に非常に類似した特徴を持つ細胞株であることが判明した。本細胞株に対してカチオン性脂質を用いた遺伝子導入でも高い導入効率を示し, 分子生物学的手法を用いた研究においても非常に有用であった。
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.

研究種目 : 基盤研究(C)(一般)
研究期間 : 2015～2017
課題番号 : 15K10420
研究分野 : 運動器