Chordoma-derived cell line U-CH1-N recapitulates the biological properties of notochordal nucleus pulposus cells

Nobuyuki Fujita, Satoshi Suzuki, Kota Watanabe, Ken Ishii, Ryuichi Watanabe, Masayuki Shimoda, Keiyo Takubo, Takashi Tsuji, Yoshiaki Toyama, Takeshi Miyamoto, Keisuke Horiuchi, Masaya Nakamura, Morio Matsumoto

Research output: Contribution to journalArticle

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Abstract

Intervertebral disc degeneration proceeds with age and is one of the major causes of lumbar pain and degenerative lumbar spine diseases. However, studies in the field of intervertebral disc biology have been hampered by the lack of reliable cell lines that can be used for in vitro assays. In this study, we show that a chordoma-derived cell line U-CH1-N cells highly express the nucleus pulposus (NP) marker genes, including T (encodes T brachyury transcription factor), 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 extracellular matrix (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 and that T plays crucial roles in maintaining the notochordal NP cell-like phenotype in this cell line. Taken together, our data indicate that U-CH1-N may serve as a useful tool in studying the biology of intervertebral disc.

Original languageEnglish
Pages (from-to)1341-1350
Number of pages10
JournalJournal of Orthopaedic Research
Volume34
Issue number8
DOIs
Publication statusPublished - 01-08-2016

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Chordoma
Cell Line
Intervertebral Disc
Extracellular Matrix
Genes
Chondrogenesis
Intervertebral Disc Degeneration
Alcian Blue
Extracellular Matrix Proteins
Essential Genes
Proliferating Cell Nuclear Antigen
Nucleus Pulposus
DNA Replication
Flow Cytometry
Spine
Transcription Factors
Immunohistochemistry
Staining and Labeling
Phenotype
Pain

All Science Journal Classification (ASJC) codes

  • Orthopedics and Sports Medicine

Cite this

Fujita, N., Suzuki, S., Watanabe, K., Ishii, K., Watanabe, R., Shimoda, M., ... Matsumoto, M. (2016). Chordoma-derived cell line U-CH1-N recapitulates the biological properties of notochordal nucleus pulposus cells. Journal of Orthopaedic Research, 34(8), 1341-1350. https://doi.org/10.1002/jor.23320
Fujita, Nobuyuki ; Suzuki, Satoshi ; Watanabe, Kota ; Ishii, Ken ; Watanabe, Ryuichi ; Shimoda, Masayuki ; Takubo, Keiyo ; Tsuji, Takashi ; Toyama, Yoshiaki ; Miyamoto, Takeshi ; Horiuchi, Keisuke ; Nakamura, Masaya ; Matsumoto, Morio. / Chordoma-derived cell line U-CH1-N recapitulates the biological properties of notochordal nucleus pulposus cells. In: Journal of Orthopaedic Research. 2016 ; Vol. 34, No. 8. pp. 1341-1350.
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Fujita, N, Suzuki, S, Watanabe, K, Ishii, K, Watanabe, R, Shimoda, M, Takubo, K, Tsuji, T, Toyama, Y, Miyamoto, T, Horiuchi, K, Nakamura, M & Matsumoto, M 2016, 'Chordoma-derived cell line U-CH1-N recapitulates the biological properties of notochordal nucleus pulposus cells', Journal of Orthopaedic Research, vol. 34, no. 8, pp. 1341-1350. https://doi.org/10.1002/jor.23320

Chordoma-derived cell line U-CH1-N recapitulates the biological properties of notochordal nucleus pulposus cells. / Fujita, Nobuyuki; Suzuki, Satoshi; Watanabe, Kota; Ishii, Ken; Watanabe, Ryuichi; Shimoda, Masayuki; Takubo, Keiyo; Tsuji, Takashi; Toyama, Yoshiaki; Miyamoto, Takeshi; Horiuchi, Keisuke; Nakamura, Masaya; Matsumoto, Morio.

In: Journal of Orthopaedic Research, Vol. 34, No. 8, 01.08.2016, p. 1341-1350.

Research output: Contribution to journalArticle

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T1 - Chordoma-derived cell line U-CH1-N recapitulates the biological properties of notochordal nucleus pulposus cells

AU - Fujita, Nobuyuki

AU - Suzuki, Satoshi

AU - Watanabe, Kota

AU - Ishii, Ken

AU - Watanabe, Ryuichi

AU - Shimoda, Masayuki

AU - Takubo, Keiyo

AU - Tsuji, Takashi

AU - Toyama, Yoshiaki

AU - Miyamoto, Takeshi

AU - Horiuchi, Keisuke

AU - Nakamura, Masaya

AU - Matsumoto, Morio

PY - 2016/8/1

Y1 - 2016/8/1

N2 - Intervertebral disc degeneration proceeds with age and is one of the major causes of lumbar pain and degenerative lumbar spine diseases. However, studies in the field of intervertebral disc biology have been hampered by the lack of reliable cell lines that can be used for in vitro assays. In this study, we show that a chordoma-derived cell line U-CH1-N cells highly express the nucleus pulposus (NP) marker genes, including T (encodes T brachyury transcription factor), 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 extracellular matrix (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 and that T plays crucial roles in maintaining the notochordal NP cell-like phenotype in this cell line. Taken together, our data indicate that U-CH1-N may serve as a useful tool in studying the biology of intervertebral disc.

AB - Intervertebral disc degeneration proceeds with age and is one of the major causes of lumbar pain and degenerative lumbar spine diseases. However, studies in the field of intervertebral disc biology have been hampered by the lack of reliable cell lines that can be used for in vitro assays. In this study, we show that a chordoma-derived cell line U-CH1-N cells highly express the nucleus pulposus (NP) marker genes, including T (encodes T brachyury transcription factor), 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 extracellular matrix (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 and that T plays crucial roles in maintaining the notochordal NP cell-like phenotype in this cell line. Taken together, our data indicate that U-CH1-N may serve as a useful tool in studying the biology of intervertebral disc.

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