Changes in the Molecular Phenotype of Nucleus Pulposus Cells with Intervertebral Disc Aging

Tang, Xinyan; Jing, Liufang; Chen, Jun

doi:10.1371/journal.pone.0052020

Cited by 90 publications

(129 citation statements)

References 76 publications

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“…Microarrays (comparison between 1 and 4-month-old NP and AF) [76] Embryonic transcription factor regulating the transcription of genes required for mesoderm formation and differentiation…”

Section: T (Brachyury) Ratmentioning

confidence: 99%

“…For this to be possible, a comprehensive phenotypic characterization of notochordal cells, and especially of markers specific to the notochordal cells in the human IVD would be required. To date, several attempts have been made to identify notochordal markers, either by comparing the gene expression of immature notochordal rat NP with its costal cartilage [73], rat NP tissue with different degrees of maturity [74][75][76], comparing immature pig NP with AF cells [74,77], sorted large and granular with smaller, less granular pig NP cells [78], immature human NP with AF cells [74] or NP from human juvenile scoliotic and adult discs [79] (Tables 1, 2, 3, 4 list putative notochordal/ immature NP markers identified in different species and describe their relevance to the IVD field). Proposed rat notochordal markers identified in these studies were CD55 [73], brachyury, neuropilin (Nrp-1), CD221, BASP-1 [76], N-Cad [73,76], TGF-b, BMP-6 and CTGF [75].…”

Section: T (Brachyury) Ratmentioning

confidence: 99%

“…Proposed human young markers were CD24, CD54 and brachyury [79]. CD90 was not expressed either in immature rat [76] or in scoliotic juvenile NP [79] and has been proposed as a negative notochordal cell marker. Limitations to these studies are the use of animal models, which may not be representative of the human intervertebral disc phenotype and the use of scoliotic discs as a source of human immature NP, as the scoliotic IVD, although derived from young patients, is prematurely degenerated and subjected In cultured IVD cells, its expression is up-regulated by the exposure to IL-17, IFNc, and TNFa [87] CD55/DAF (decay accelerating factor for complement)…”

Section: T (Brachyury) Ratmentioning

confidence: 99%

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An understanding of intervertebral disc development, maturation and cell phenotype provides clues to direct cell-based tissue regeneration therapies for disc degeneration

2014

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Cell-based regenerative medicine therapies have been proposed for repairing the degenerated intervertebral disc (a major cause of back pain). However, for this approach to be successful, it is essential to characterise the phenotype of its native cells to guarantee that implanted cells differentiate and maintain the correct phenotype to ensure appropriate cell and tissue function. While recent studies have increased our knowledge of the human nucleus pulposus (NP) cell phenotype, their ontogeny is still unclear. The expression of notochordal markers by a subpopulation of adult NP cells suggests that, contrary to previous reports, notochord-derived cells are retained in the adult NP, possibly coexisting with a second population of cells originating from the annulus fibrosus or endplate. It is not known, however, how these two cell populations interact and their specific role(s) in disc homeostasis and disease. In particular, notochordal cells are proposed to display both anabolic and protective roles; therefore, they may be the ideal cells to repair the degenerate disc. Thus, understanding the ontogeny of the adult NP cells is paramount, as it will inform the medical and scientific communities as to the ideal phenotype to implant into the degenerate disc and the specific pathways involved in stem cell differentiation towards such a phenotype.

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“…Microarrays (comparison between 1 and 4-month-old NP and AF) [76] Embryonic transcription factor regulating the transcription of genes required for mesoderm formation and differentiation…”

Section: T (Brachyury) Ratmentioning

confidence: 99%

Section: T (Brachyury) Ratmentioning

confidence: 99%

Section: T (Brachyury) Ratmentioning

confidence: 99%

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An understanding of intervertebral disc development, maturation and cell phenotype provides clues to direct cell-based tissue regeneration therapies for disc degeneration

2014

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“…Les cellules iPS -qui, contrairement aux CSE, ne posent pas les problèmes éthiques d'utilisation d'embryonspourraient constituer un tournant dans la stratégie de prise en charge de la dégénérescence discale. Ainsi, il a été récemment montré que des cellules iPS murines, différenciées en corps embryoïdes, dont les cellules sont triées selon leur expression de CD24, un marqueur de cellules progénitrices du NP [22], pouvaient se différencier en une population cellulaire proche des nucléopulpocytes. Ces cellules se sont révélées capables de synthétiser les composants de la MEC d'un NP natif (protéoglycanes et collagène de type II) [23].…”

Section: Synthèse Revuesunclassified

Médecine régénératrice du disque intervertébral

et al. 2014

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“…For each sample in microarray, tissue was pooled from all discs across four rats to collect sufficient RNA for one sample [59]. The amount and quality of RNA used was not mentioned in their study.…”

Section: Yield Of Rna From Rat Np Tissuementioning

confidence: 99%

Technique and method development for intervertebral disc (IVD) research

Lee¹,

李芷恩²

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Intervertebral disc (IVD) degeneration is one of the major causes of low back pain. The direct and indirect cost of managing back pain posts heavy socioeconomic burden to the society. Improved technologies and techniques together with well documented experiments will benefit the research field by saving effort in doing the optimization in every laboratory and avoiding experiment failure due to incomplete understanding of the procedures. These can accelerate scientific discovery, reduce the sacrifice of animals and enable a more effective use of funding.Nucleus pulposus (NP) is the central part of IVD. Differences in matrix compositions in human NP clinical samples demand different cell isolation protocols for optimal results but there is no clear guide about this to date. Sub-optimal protocols may result in low cell yield, limited reliability of results or even failure of experiments. We experimented different isolation protocols to study different parameters involved and suggested some rules for cell isolation in three main applications: RNA extraction for phenotyping, cell isolation for cell culture, and characterization by flow cytometry.In addition, instead of extracting RNA from isolated cells, extraction of RNA from tissues directly may avoid the change of RNA levels during the cell isolation process.However, extraction of RNA directly from human and large animal IVD tissue is technically challenging due to its tough nature, low cell-to-matrix ratio and high proteoglycan content. Thus we developed a method for RNA extraction from bovine disc tissues by integrating the use of cryosectioning, additional phase separation and high salt precipitation into conventional guanidinium thiocyanate based method. With this method, RNA could be extracted from the NP tissue directly but the concentration was low. A shift toward 270 nm was observed in its UV spectrum which was due to phenol contamination. This caused an overestimation of RNA concentration. Hence we developed a computational method based on UV spectra for correcting the In short, different methods related to IVD research were developed and optimized. With improved methods together with a better understanding of the underlying rationale, researchers can save time and cost in their experiments and reduce experiment failure rate. This will help to accelerate researches. New methods also enable studies which were not feasible in the past.(484 words)

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Changes in the Molecular Phenotype of Nucleus Pulposus Cells with Intervertebral Disc Aging

Cited by 90 publications

References 76 publications

An understanding of intervertebral disc development, maturation and cell phenotype provides clues to direct cell-based tissue regeneration therapies for disc degeneration

An understanding of intervertebral disc development, maturation and cell phenotype provides clues to direct cell-based tissue regeneration therapies for disc degeneration

Médecine régénératrice du disque intervertébral

Technique and method development for intervertebral disc (IVD) research

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