Same-species phenotypic comparison of notochordal and mature nucleus pulposus cells

Saggese, Taryn; Redey, Prutha; McGlashan, Susan R.

doi:10.1007/s00586-014-3697-9

Cited by 12 publications

(25 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…T is an embryonic transcription factor required for mesoderm formation and differentiation and notochord‐development . This protein has often been used as a marker of a notochordal phenotype . In our study, T was found to be expressed by all notochordal cells at all stages analyzed, which suggests that this transcription factor has a role in human notochordal cell development.…”

Section: Discussionsupporting

confidence: 61%

Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord‐specific markers during early human intervertebral disc development

Rodrigues-Pinto

Berry

Hanley

et al. 2016

Journal Orthopaedic Research

View full text Add to dashboard Cite

In humans, the nucleus pulposus (NP) is composed of large vacuolated notochordal cells in the fetus but, soon after birth, becomes populated by smaller, chondrocyte‐like cells. Although animal studies indicate that notochord‐derived cells persist in the adult NP, the ontogeny of the adult human NP cell population is still unclear. As such, identification of unique notochordal markers is required. This study was conducted to determine the spatiotemporal expression of putative human notochordal markers to aid in the elucidation of the ontogeny of adult human NP cells. Human embryos and fetuses (3.5–18 weeks post‐conception (WPC)) were microdissected to isolate the spine anlagens (notochord and somites/sclerotome). Morphology of the developing IVD was assessed using hematoxylin and eosin. Expression of keratin (KRT) 8, KRT18, KRT19, CD24, GAL3, CD55, BASP1, CTGF, T, CD90, Tie2, and E‐cadherin was assessed using immunohistochemistry. KRT8, KRT18, KRT19 were uniquely expressed by notochordal cells at all spine levels at all stages studied; CD24 was expressed at all stages except 3.5 WPC. While GAL3, CD55, BASP1, CTGF, and T were expressed by notochordal cells at specific stages, they were also co‐expressed by sclerotomal cells. CD90, Tie2, and E‐cadherin expression was not detectable in developing human spine cells at any stage. This study has identified, for the first time, the consistent expression of KRT8, KRT18, KRT19, and CD24 as human notochord‐specific markers during early IVD development. Thus, we propose that these markers can be used to help ascertain the ontogeny of adult human NP cells. © 2016 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. J Orthop Res 34:1327–1340, 2016.

show abstract

Section: Discussionsupporting

confidence: 61%

Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord‐specific markers during early human intervertebral disc development

Rodrigues-Pinto

Berry

Hanley

et al. 2016

Journal Orthopaedic Research

View full text Add to dashboard Cite

show abstract

“…Due to the broad donor age range used in this study (1-8.5 years old), NP tissue may contain varying amounts of notochordal cells (NC). It has been suggested that up to 10% of bovine NP cells per tail IVD are NC, based on size exclusion and/or Keratin 8 expression [17,24]. The latter study used 18-24 month old bovine donors, while the former study did not report donor age.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, model organisms, such as the cow, are used for in vitro cellular and ex vivo biomechanical studies of the IVD [14,15]. The bovine tail IVD represents a good model for the non-degenerate human IVD: the bovine NP retains few notochordal cells and whole transcriptome analyses identified similar phenotypic marker genes as in humans [6,16,17]. This study aimed to establish whether IVD phenotypic marker genes, which we previously used to distinguish non-degenerate human NP from AF cells, can be used to describe the cell phenotype in the outer AF, inner AF and NP in situ in the bovine tail IVD and this was compared to passage 0 monolayer cells.…”

Section: Introductionmentioning

confidence: 99%

Transcriptional profiling distinguishes inner and outer annulus fibrosus from nucleus pulposus in the bovine intervertebral disc

et al. 2017

View full text Add to dashboard Cite

Background Cells in the intervertebral disc have unique phenotypes and marker genes that separate the nucleus pulposus (NP), annulus fibrosus (AF) and articular cartilage (AC) have been identified. Recently, it was shown that phenotypic marker genes exhibit variable expression in humans. In this study, the bovine tail was used to determine the ability of marker genes to distinguish the outer and inner AF from NP tissue and isolated cells. Methods Bovine tail intervertebral discs from 13 donors were dissected and correct isolation of tissue was confirmed. mRNA was isolated directly from tissue or passage 0 monolayer cells and used for gene expression measurements (qPCR). Conventional marker genes (bAcan, bCol1a1, bCol2a1) and novel marker genes (bAdamts17, bBrachyury/T, bCD24, bCol5a1, bCol12a1, bFoxf1, bKrt19, bPax1, bSfrp2) were evaluated. Results As expected bAcan, bCol2a1 and bCol1a1 distinguished outer AF from NP tissue, while inner AF and NP could not be discriminated. The NP markers bT, bCd24 and bKrt19 were significantly higher expressed in NP than inner and outer AF tissue. bFoxF1 and bPax1 only distinguished IVD tissues from AC. The AF markers bAdamts17, bCol5a1, bCol12a1 and bSfrp2 were higher expressed in the outer AF compared with inner AF and NP tissue. Monolayer culturing strongly decreased bAcan, bCol2a1, bCD24 and bCol5a1 expression, while bCol1a1, bT, bKrt19 and bSfrp2 were not affected. ConclusionThe IVD phenotypic marker genes bT, bKrt19, bSfrp2 and bCol12a1 convincingly distinguished NP from outer AF in situ and in vitro.

show abstract

“…NC-conditioned medium (NCCM, containing factors secreted by NCs) and NC:CLC co-culture stimulate in vitro differentiation of MSCs into a NP-like phenotype Korecki et al, 2010;Purmessur et al, 2011), protect the NP from apoptosis (Erwin et al, 2011;Mehrkens et al, 2013), and/or increase CLC proliferation and GAG production (Abbott et al, 2012;Aguiar et al, 1999;Gantenbein et al, 2014;Potier et al, 2014). In addition, NCs significantly stimulate CLC activity and GAG production (Gantenbein-Ritter et al, 2012) and produce more GAGs than CLCs (Saggese et al, 2014). NCs have also been described to exert potential symptom modifying actions, e.g.…”

Section: Ncs Of Different Species Regenerate Human Clcsmentioning

confidence: 99%

The species-specific regenerative effects of notochordal cell-conditioned medium on chondrocyte-like cells derived from degenerated human intervertebral discs

Bach

Vries

Krouwels

et al. 2015

eCM

View full text Add to dashboard Cite

During intervertebral disc (IVD) maturation, the main cell type shifts from notochordal cells (NCs) to chondrocytelike cells (CLCs). NCs secrete factors with regenerative potential, making them an interesting focus for regenerative treatments. During initial development, these strategies preferably employ non-human donors due to easy availability of their NC-rich nucleus pulposus (NP) tissue. To increase the success of translating these strategies for clinical application, this study aimed to delineate whether NC-secreted factors of different species have a regenerative effect on human CLCs. Human, canine and porcine NC-rich NP tissue and NC-conditioned medium (NCCM) were analysed biochemically and histologically. Human CLC micro-aggregates from degenerated IVDs were cultured in human, canine or porcine NCCM. Collagen, glycosaminoglycan (GAG) and DNA content was determined and histology was performed. Canine and porcine NPs were richer in NCs than human NPs. Human NPs contained the highest collagen content, whereas the DNA and GAG content of canine NPs was significantly higher than that of human or porcine NPs. NCCM from all species significantly increased the DNA and GAG content of the human CLC micro-aggregates. Porcine and canine NCCM were significantly more potent than human NCCM in inducing GAG deposition, whereas only human NCCM induced collagen type II production. Secreted factors from human, canine and porcine NC-rich NPs exerted regenerative effects on human CLCs, indicating a cross-species effect. Bioactive compound(s) are present in NCCM of different species that may reverse human IVD degeneration, supporting further research into strategies based on NC-technology employing canine or porcine models for their translation into humans.

show abstract

Same-species phenotypic comparison of notochordal and mature nucleus pulposus cells

Abstract: NC and MNP cells can be isolated from the same bovine disc and maintain their distinct phenotypes in 3D culture.

Cited by 12 publications

References 28 publications

Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord‐specific markers during early human intervertebral disc development

Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord‐specific markers during early human intervertebral disc development

Transcriptional profiling distinguishes inner and outer annulus fibrosus from nucleus pulposus in the bovine intervertebral disc

The species-specific regenerative effects of notochordal cell-conditioned medium on chondrocyte-like cells derived from degenerated human intervertebral discs

Contact Info

Product

Resources

About