Nucleus pulposus phenotypic markers to determine stem cell differentiation: fact or fiction?

Thorpe, A; Binch, Abbie L. A.; Creemers, Laura B.; Sammon, Chris; Maitre, Christine L. Le

doi:10.18632/oncotarget.6782

Cited by 59 publications

(66 citation statements)

References 53 publications

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“…Compared to conventional marker genes bT and bKRT19 were rather difficult to detect, exemplified by the large number of non-detectable samples in tissues with low expression. In agreement with other studies, we found that the NP/IVD can be distinguished from AC by high expression of bPax1 and bFoxf1 [7,13]. These genes exhibited the highest differential expression in our study.…”

Section: Discussionsupporting

confidence: 93%

“…In addition, NP specific marker genes are increasingly used as readout to develop stem cell differentiation protocols for NP regeneration [11,12]. However, a recent report described large variation for NP and AF marker genes in human cell isolates, which prevented separation of NP and AF tissue based on phenotypic markers [13]. Only PAX1 and FOXF1 were confirmed to distinguish NP tissue from AC and at the protein level NP from AC.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2017

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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.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

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

et al. 2017

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“…As expected, the knockout of all WNT genes led to significantly decreased PAX1 and FOXF1 expression, markers suggested to be representative of the healthy, mature NP cell phenotype . Interestingly, the overexpression of WNT5A and WNT11 genes increased PAX1 expression by five‐ and ninefold, respectively, versus the GFP control at day 0 of redifferentiation; however, this effect was attenuated by day 28.…”

Section: Discussionsupporting

confidence: 64%

Impact of Wnt signals on human intervertebral disc cell regeneration

Pizzute

Zhang

et al. 2018

Journal Orthopaedic Research

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Although preconditioning strategies are growing areas of interest for therapies targeting intervertebral discs (IVDs), it is unknown whether the Wnt signals previously implicated in chondrogenesis, Wnt3A, Wnt5A, and Wnt11, play key roles in the promotion of human nucleus pulposus (NP) cell redifferentiation. In this study, NP cells isolated from herniated disc patients were transduced with lentiviral vectors to overexpress the WNT3A, WNT5A, or WNT11 genes, or CRISPR associated protein 9 (Cas9)/single-guide RNA (sgRNA) vectors to knock out these genes. Following expansion, transduced NP cells were induced for redifferentiation toward the NP phenotype. The overexpression of specific WNT factors led to increases in both glycosaminoglycan (GAG) deposition and expression of redifferentiation genes. These effects were attenuated by knockout of the same WNT genes. These results indicate that specific WNT signals can regulate the expression of redifferentiation genes, unequally impact GAG deposition, and contribute to the redifferentiation of human NP cells. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

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“…Several previous studies have investigated specific markers for NP cells [42-45]. In this study, high-magnitude (20% deformation) compression down-regulated the expression of these NP markers (keratin 19, glypican and CA XII) compared with low-magnitude (2% deformation) compression.…”

Section: Discussionmentioning

confidence: 66%

N-Cadherin Maintains the Healthy Biology of Nucleus Pulposus Cells under High-Magnitude Compression

Wang

Leng²,

Zhao³

et al. 2017

Cell Physiol Biochem

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Background/Aims: Mechanical load can regulate disc nucleus pulposus (NP) biology in terms of cell viability, matrix homeostasis and cell phenotype. N-cadherin (N-CDH) is a molecular marker of NP cells. This study investigated the role of N-CDH in maintaining NP cell phenotype, NP matrix synthesis and NP cell viability under high-magnitude compression. Methods: Rat NP cells seeded on scaffolds were perfusion-cultured using a self-developed perfusion bioreactor for 5 days. NP cell biology in terms of cell apoptosis, matrix biosynthesis and cell phenotype was studied after the cells were subjected to different compressive magnitudes (low- and high-magnitudes: 2% and 20% compressive deformation, respectively). Non-loaded NP cells were used as controls. Lentivirus-mediated N-CDH overexpression was used to further investigate the role of N-CDH under high-magnitude compression. Results: The 20% deformation compression condition significantly decreased N-CDH expression compared with the 2% deformation compression and control conditions. Meanwhile, 20% deformation compression increased the number of apoptotic NP cells, up-regulated the expression of Bax and cleaved-caspase-3 and down-regulated the expression of Bcl-2, matrix macromolecules (aggrecan and collagen II) and NP cell markers (glypican-3, CAXII and keratin-19) compared with 2% deformation compression. Additionally, N-CDH overexpression attenuated the effects of 20% deformation compression on NP cell biology in relation to the designated parameters. Conclusion: N-CDH helps to restore the cell viability, matrix biosynthesis and cellular phenotype of NP cells under high-magnitude compression.

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Nucleus pulposus phenotypic markers to determine stem cell differentiation: fact or fiction?

Cited by 59 publications

References 53 publications

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

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

Impact of Wnt signals on human intervertebral disc cell regeneration

N-Cadherin Maintains the Healthy Biology of Nucleus Pulposus Cells under High-Magnitude Compression

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