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

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. © 2016 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 34:1341–1350, 2016.

Melatonin reverses tumor necrosis factor-alpha-induced metabolic disturbance of human nucleus pulposus cells via MTNR1B/Gαi2/YAP signaling

“…Interestingly, our previous study showed that MTNR1B can also interact with Gαi2 in the chordoma cell lines, U-CH1 and MUG-CC1 [54]. We speculate that this coincidence may be related to the homology between NP cells and chordoma cell lines [57].…”

Efficacy of hyaluronic acid on intervertebral disc inflammation: An in vitro study using notochordal cell lines and human disc cells

“…U-CH1-N cells are a chondroma-derived cell line that share certain biological characteristics with notochordal NP cells. 33 Cells were cultured in minimum essential medium alpha (MEM-alpha; Sigma-Aldrich) with 10% FBS and 1% penicillin-streptomycin in a humidified atmosphere containing 5% CO 2 at 37°C. U-CH1-N cells were seeded into six-well plates at a density of 2 × 10 5 cells/well or into 100 mm dishes at a density of 1 × 10 6 cells/dish 1 day before treatment, respectively.…”

New Horizons in Spine Research: Disc biology, spine biomechanics and pathomechanisms of back pain

“…This review follows the consensus review on nucleus pulposus phenotype of the ORS Spine Research Interest Group (now Spine Section) . Improved functional nucleus pulposus cell markers are reported that highlight notochordal markers, clarify their distinction from articular chondrocytes, and include novel molecular phenotypic changes that also occur with ageing and degeneration . Defining markers of annulus fibrosus phenotype remains a young field and important functional cellular behaviours of annulus cells are described …”