Novel Immortal Cell Lines Support Cellular Heterogeneity in the Human Annulus Fibrosus

PloS One
Guus G H van den AkkerTim J M Welting

Abstract

Loss of annulus fibrosus (AF) integrity predisposes to disc herniation and is associated with IVD degeneration. Successful implementation of biomedical intervention therapy requires in-depth knowledge of IVD cell biology. We recently generated unique clonal human nucleus pulposus (NP) cell lines. Recurring functional cellular phenotypes from independent donors provided pivotal evidence for cell heterogeneity in the mature human NP. In this study we aimed to generate and characterize immortal cell lines for the human AF from matched donors. Non-degenerate healthy disc material was obtained as surplus surgical material. AF cells were immortalized by simian virus Large T antigen (SV40LTAg) and human telomerase (hTERT) expression. Early passage cells and immortalized cell clones were characterized based on marker gene expression under standardized culturing and in the presence of Transforming Growth factor β (TGFβ). The AF-specific expression signature included COL1A1, COL5A1, COL12A1, SFRP2 and was largely maintained in immortal AF cell lines. Remarkably, TGFβ induced rapid 3D sheet formation in a subgroup of AF clones. This phenotype was associated with inherent differences in Procollagen type I processing and maturation, and cor...Continue Reading

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Citations

Mar 16, 2016·BMC Musculoskeletal Disorders·Guus G H van den AkkerJan Willem Voncken
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Aug 24, 2018·Journal of Orthopaedic Surgery and Research·Ann-Kathrin SchubertJochen Ringe
Apr 21, 2021·Aerospace Medicine and Human Performance·Karenna Smith, Jeremy Mercuri
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Nov 30, 2021·Journal of Biomedical Materials Research. Part B, Applied Biomaterials·Ryan BoremJeremy Mercuri

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Methods Mentioned

BETA
biopsies
PCR
protein assay

Software Mentioned

Pannoramic Viewer
DeltaPix

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