Matrix protein gene expression in intervertebral disc cells subjected to altered osmolarity

Chen, Jun; Baer, Anthony E.; Paik, Phil; Yan, Wei; Setton, Lori A.

doi:10.1016/s0006-291x(02)00314-5

Cited by 94 publications

(99 citation statements)

References 41 publications

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“…There are a number of studies that investigated the influence of hydrostatic and osmotic pressure on disc tissue. Chen et al showed that under hypo-osmotic (255 mOsm) conditions in vitro gene expression of aggrecan and collagen type II was upregulated in the transition zone of the disc [3]. In contrast, expression within the nucleus pulposus was not significantly changed.…”

Section: Discussionmentioning

confidence: 99%

Intradiscal pressure measurements in normal discs, compressed discs and compressed discs treated with axial posterior disc distraction: an experimental study on the rabbit lumbar spine model

et al. 2005

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Section: Discussionmentioning

confidence: 99%

Intradiscal pressure measurements in normal discs, compressed discs and compressed discs treated with axial posterior disc distraction: an experimental study on the rabbit lumbar spine model

et al. 2005

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“…Cells were isolated from tissue harvested from the NP and AF of spines with a sequential pronase-collagenase digestion [7]. The primary cells were suspended in cell culture media (Ham's F-12 medium, Invitrogen Life Technologies, Carlsbad, CA, USA; supplemented with 10% fetal bovine serum (FBS), 25 lg/ml ascorbic acid, 100 U/ml penicillin, 100 lg/ml streptomycin, 1 lg/ml fungizone and 10 lg/ml DNase I) prior to cell sorting.…”

Section: Primary Cell Isolationmentioning

confidence: 99%

“…Quantification of mRNA was performed for genes of a wide panel of proteins relevant to disc homeostasis, including key extracellular matrix proteins, metalloproteinases and their inhibitors, cytoskeletal proteins, cytokines, growth factors, stress response proteins and transcriptional factors. For each target gene, two porcine-specific PCR primers and one fluorescently labeled intron-spanning probe were designed using published sequences in GenBank and Primer Expressä software (Applied Biosystems, Foster City, CA, see Table 1 and Chen et al 7 for sequence details). Real time RT-PCR (SmartCycler Ò system, Cepheid, Sunnyvale, CA, USA) conditions were used as described previously [7].…”

Section: Primary Cell Isolationmentioning

confidence: 99%

“…For each target gene, two porcine-specific PCR primers and one fluorescently labeled intron-spanning probe were designed using published sequences in GenBank and Primer Expressä software (Applied Biosystems, Foster City, CA, see Table 1 and Chen et al 7 for sequence details). Real time RT-PCR (SmartCycler Ò system, Cepheid, Sunnyvale, CA, USA) conditions were used as described previously [7]. Relative gene expression differences were quantified among the sorted notochordal-like cells, small NP cells and anulus fibrosus cells using the comparative Ct method with 18S rRNA as an internal control [7].…”

Section: Primary Cell Isolationmentioning

confidence: 99%

“…Real time RT-PCR (SmartCycler Ò system, Cepheid, Sunnyvale, CA, USA) conditions were used as described previously [7]. Relative gene expression differences were quantified among the sorted notochordal-like cells, small NP cells and anulus fibrosus cells using the comparative Ct method with 18S rRNA as an internal control [7]. Duplicate PCR reactions were performed for each target gene and the internal control for one RNA sample.…”

Section: Primary Cell Isolationmentioning

confidence: 99%

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Molecular phenotypes of notochordal cells purified from immature nucleus pulposus

2006

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The immature nucleus pulposus (NP) is populated by cells of notochordal-origin that are larger and contain an extensive cytoskeletal network and numerous vacuoles. The disappearance of these cells with age is believed important in regulating metabolic shifts that may contribute to age-related disc degeneration. The precise biological function of these notochordal cells in the immature NP remains unclear, however, because of challenges in studying the mixed cell population in the NP. In this study, notochordallike cells were purified from immature NP cells using a new fluorescence-activated cell sorting (FACS) protocol with auto-fluorescence and size analysis. The unique molecular phenotypes of sorted notochordallike cells were characterized by the mRNA expression pattern for key matrix proteins and modulators, and by the expression of cell-matrix receptor integrin subunits. An FACS analysis showed that the immature NP contained a majority of cells that were larger than anulus fibrosus (AF) cells and with fluorescence higher than AF cells. In comparison with the small NP cells separated by the FACS protocol, sorted notochordal-like cells expressed lower mRNA levels of type I collagen, biglycan, TIMP1, HSP70 and c-fos, and did not express detectable mRNA levels of decorin, lumican, multiple MMPs or IL-1b via realtime quantitative RT-PCR. A greater number of these notochordal-like cells also expressed the higher levels of a6, a1 and b1 integrin subunits as compared to small NP cells. Together, our results point towards a unique molecular phenotype for these notochordal-like cells of NP, characterized by the absence of gene expression for specific small proteoglycans and higher protein expression of integrin subunits that regulate interactions with collagens and laminin. Future studies will be important for revealing if this unique molecular profile is coordinated with functional differences in pericellular matrix regions and/or integrin-mediated cell-matrix interactions for these notochordallike cells within the NP.

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Surgical anatomy, radiological features, and molecular biology of the lumbar intervertebral discs

et al. 2017

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The intervertebral disc (IVD) is a joint unique in structure and functions. Lying between adjacent vertebrae, it provides both the primary support and the elasticity required for the spine to move stably. Various aspects of the IVD have long been studied by researchers seeking a better understanding of its dynamics, aging, and subsequent disorders. In this article, we review the surgical anatomy, imaging modalities, and molecular biology of the lumbar IVD. Clin. Anat. 30:251-266, 2017. © 2017 Wiley Periodicals, Inc.

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Matrix protein gene expression in intervertebral disc cells subjected to altered osmolarity

Cited by 94 publications

References 41 publications

Intradiscal pressure measurements in normal discs, compressed discs and compressed discs treated with axial posterior disc distraction: an experimental study on the rabbit lumbar spine model

Intradiscal pressure measurements in normal discs, compressed discs and compressed discs treated with axial posterior disc distraction: an experimental study on the rabbit lumbar spine model

Molecular phenotypes of notochordal cells purified from immature nucleus pulposus

Surgical anatomy, radiological features, and molecular biology of the lumbar intervertebral discs

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