The evolutionary importance of cell ratio between notochordal and nucleus pulposus cells: an experimental 3-D co-culture study

Gantenbein, Benjamin; Chan, Samantha

doi:10.1007/s00586-011-2026-9

Cited by 37 publications

(42 citation statements)

References 33 publications

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“…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

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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.

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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

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“…It may be readily combined with cells to form a three-dimensional hydrogel structure under mild crosslinking conditions using calcium chloride (CaCl2). Alginate has been shown to support a wide variety of cell types including NP cells (29)(30)(31) and MSCs (32,33).…”

Section: Introductionmentioning

confidence: 99%

Differential Response of Encapsulated Nucleus Pulposus and Bone Marrow Stem Cells in Isolation and Coculture in Alginate and Chitosan Hydrogels

Naqvi

Buckley

2015

Tissue Engineering Part A

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Cell-based therapies may hold significant promise for the treatment of early stage degeneration of the intervertebral disc (IVD). Given their propensity to proliferate and ability to form multiple tissue types, mesenchymal stem cells (MSCs) have been proposed as a potential cell source to promote repair of the nucleus pulposus (NP). However, for any successful cell-based therapy a carrier biomaterial may be essential for targeted delivery providing key biophysical and biochemical cues to facilitate differentiation of MSCs. Two widely used biomaterials for NP regeneration are chitosan and alginate. The primary objective of this study was to assess the influence of alginate and chitosan hydrogels on bone marrow (BM) MSCs and NP cells in isolation or in co-culture. A secondary objective of this study was to investigate co-culture seeding density effects of BM and NP cells and simultaneously explore which cell type is responsible for matrix formation in a co-cultured environment. Porcine NP and BM cells were encapsulated in alginate and chitosan hydrogels separately at two seeding densities (4x10 6 and 8x10 6 cells/ml) or in co-culture (1:1, 8 x10 6 cells/ml). Constructs (diameter:5mm, height:3mm) were maintained under IVD-like conditions (low-glucose, low (5%) oxygen) with or without TGF-β3 supplementation for 21 days. Results demonstrated differential viability depending on hydrogel type. NP cells remained viable in both biomaterial types whereas BM viability was diminished in chitosan.Furthermore, hydrogel type was found to regulate sGAG and collagen accumulation.Specifically, alginate better supports sGAG accumulation and collagen type II deposition for both NP and BM cell types compared to chitosan. Having identified that alginate more readily supports cell viability and matrix accumulation we further explored additional effects of seeding density ratios (NP: BM -1:1, 1:2) for co-culture studies. Interestingly, in coculture conditions, the BM cell population declined in number while NP cells increased indicating that MSCs may in fact be signalling NP cells to proliferate rather than contributing to matrix formation. These findings provide exciting new insights on the potential of MSCs for NP tissue regeneration strategies.

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“…The absence of effects on the gene expression of COL2A1, ACAN, or SOX9 has been reported before on NPCs 19,20 and BMSCs 22 and thus is not unanticipated. The GAG production, on the other hand, has been shown to be stimulated by NCs in NPCs 19,20 or BMSCs. 22 This lack of substantial effects on the Mix could be explained by the use of dispersed NCs seeded in alginate to stimulate the other cells.…”

Section: Potier and Itomentioning

confidence: 85%

“…These cells attract more and more attention from the disc scientific community as they seem heavily involved in disc homeostasis and their (natural) disappearance may ultimately cause disc degeneration. 18 Most importantly here, NCs have been shown to stimulate not only NPCs [19][20][21] but also BMSCs. Purmessur et al and Korecki et al, indeed, showed that factors secreted by porcine NCs can promote the synthesis of proteoglycans (main component of disc matrix with collagen) and the gene expression of discogenic markers (aggrecan and type II collagen) by human BMSCs.…”

mentioning

confidence: 99%

Can Notochordal Cells Promote Bone Marrow Stromal Cell Potential for Nucleus Pulposus Enrichment? A SimplifiedIn VitroSystem

Potier

Ito

2014

Tissue Engineering Part A

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Bone marrow stromal cells (BMSCs) have shown promising potential to stop intervertebral disc degeneration in several animal models. In order to restore a healthy state, though, this potential should be further stimulated. Notochordal cells (NCs), influential in disc development, have been shown to stimulate BMSC differentiation, but it is unclear how this effect will translate in an environment where resident disc cells (nucleus pulposus cells [NPCs]) could also influence BMSCs. The goal of this study was, therefore, to evaluate the effects of NCs on BMSCs when cocultured with NPCs, in a simplified 3D in vitro system. Bovine BMSCs and NPCs were mixed (Mix) and seeded into alginate beads. Using culture inserts, the Mix was then cocultured with porcine NCs (alginate beads) and compared to coculture with empty beads or porcine skin fibroblasts (SFs, alginate beads). NPCs alone were also cocultured with NCs, and BMSCs alone cultured under chondrogenic conditions. The effects of coculture conditions on cell viability, matrix production (proteoglycan and collagen), and gene expression of disc markers (aggrecan, type II collagen, and SOX9) were assessed after 4 weeks of culture. The NC phenotype and gene expression profile were also analyzed. Coculture with NCs did not significantly influence cell viability, proteoglycan production, or disc marker gene expression of the Mix. When compared to NPCs, the Mix produced the same amount of proteoglycan and displayed a higher expression of disc marker, indicating a stimulation of the BMSCs (and/or NPCs) in the Mix. Additionally, during the 4 weeks of culture, the NC phenotype changed drastically (morphology, gene expression profile). These results show that NCs might not be as stimulatory for BMSCs in an NPC-rich environment, as believed from individual cultures. This absence of effects could be explained by a mild stimulation provided by (de)differentiating NCs and the costimulation of BMSCs and NPCs by each other.

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The evolutionary importance of cell ratio between notochordal and nucleus pulposus cells: an experimental 3-D co-culture study

Cited by 37 publications

References 33 publications

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

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

Differential Response of Encapsulated Nucleus Pulposus and Bone Marrow Stem Cells in Isolation and Coculture in Alginate and Chitosan Hydrogels

Can Notochordal Cells Promote Bone Marrow Stromal Cell Potential for Nucleus Pulposus Enrichment? A SimplifiedIn VitroSystem

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