Sox9 expression of alginate-encapsulated chondrocytes is stimulated by low cell density

Bernstein, Peter S.; Meng, Dong; Graupher, Sylvi; Corbeil, Denis; Gelinsky, Michael; Günther, Klaus‐Peter; Fickert, Stefan

doi:10.1002/jbm.a.34995

Cited by 5 publications

(8 citation statements)

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“…For example, overall cell density, ratios of co-cultured cell types, and addition of growth factors affect chondrogenesis and cell survival both in vitro and in vivo. 39,63,64 Encapsulation materials also affect matrix synthesis and whether cells benefit from direct cell-cell contact. 37 The cell type and configuration results from our initial study provide a template to which additional elements can be added to design a more effective NP tissue engineering construct.…”

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confidence: 99%

Effects of cell type and configuration on anabolic and catabolic activity in 3D co‐culture of mesenchymal stem cells and nucleus pulposus cells

Ouyang

Cerchiari

Tang

et al. 2016

Journal Orthopaedic Research

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Tissue engineering constructs to treat intervertebral disc degeneration must adapt to the hypoxic and inflammatory degenerative disc microenvironment. The objective of this study was to determine the effects of two key design factors, cell type and cell configuration, on the regenerative potential of nucleus pulposus cell (NPC) and mesenchymal stem cell (MSC) constructs. Anabolic and catabolic activity was quantified in constructs of varying cell type (NPCs, MSCs, and a 50:50 co-culture) and varying configuration (individual cells and micropellets). Anabolic and catabolic outcomes were both dependent on cell type. Gene expression of Agg and Col2A1, glycosaminoglycan (GAG) content, and aggrecan immunohistochemistry (IHC), were significantly higher in NPC-only and co-culture groups than in MSC-only groups, with NPC-only groups exhibiting the highest anabolic gene expression levels. However, NPC-only constructs also responded to inflammation and hypoxia with significant upregulation of catabolic genes (MMP-1, MMP-9, MMP-13, and ADAMTS-5). MSC-only groups were unaffected by degenerative media conditions, and co-culture with MSCs modulated catabolic induction of the NPCs. Culturing cells in a micropellet configuration dramatically reduced catabolic induction in co-culture and NPC-only groups. Co-culture micropellets, which take advantage of both cell type and configuration effects, had the most immunomodulatory response, with a significant decrease in MMP-13 and ADAMTS-5 expression in hypoxic and inflammatory media conditions. Co-culture micropellets were also found to self-organize into bilaminar formations with an MSC core and NPC outer layer. Further understanding of these cell type and configuration effects can improve tissue engineering designs.

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confidence: 99%

Effects of cell type and configuration on anabolic and catabolic activity in 3D co‐culture of mesenchymal stem cells and nucleus pulposus cells

Ouyang

Cerchiari

Tang

et al. 2016

Journal Orthopaedic Research

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“…22 The expression of Sox9 increased during the¯rst 2 weeks of culture and then declined thereafter, which is consistent with previous studies. 12 As an upstream regulator, the gene expression of COL II was similar to Sox9 in the 0.25, 2.5 and 25 K groups. 23 However, the expression of COL II was not proportional to Sox9 in the 250 K group.…”

Section: Discussionmentioning

confidence: 85%

“…24 This e®ect might arise from a Sox9-sensitive gene aggrecan. 12 CellÀcell contact and interaction between chondrocytes are directly regulated and controlled by cadherins. Beta-catenin is linked to cadherin-based adhesive junctions, which are in turn in°uenced by the wnt signaling pathway.…”

Section: Discussionmentioning

confidence: 99%

“…10À15 Raising cell number from 1 to 25 million cells/mL/alginate bead could enhance the production of glycosaminoglycan (GAG) at the expense of the functional phenotype of individual cells, but an extra-high cell number of 70 million cells/mL alginate may hinder chondrogenic di®erentiation within alginate beads. 10,12 In another study, 0.25 to 11 million chondrocytes were seeded onto 5 mm diameter wells to evaluate their aggregation and chondrogenesis. An optimal initial seeding number of 3.75 million cells was determined upon evaluation of gross morphology, histological, biochemical and biomechanical data.…”

Section: Introductionmentioning

confidence: 99%

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RELATIONSHIP BETWEEN CELL FUNCTION AND INITIAL CELL SEEDING DENSITY OF PRIMARY PORCINE CHONDROCYTES IN VITRO

Zhang

Wang

Han

et al. 2013

Biomed. Eng. Appl. Basis Commun.

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Maintenance of differentiated functional phenotype within in vitro chondrocyte culture requires seeding at high densities with large numbers of cells. However, optimal cell seeding numbers and densities remain elusive due to multiple varying parameters and different methodologies utilized in previous studies. In the current study, we tried to investigate the relationship between cell seeding number and differentiated functional phenotype of in vitro cultured chondrocytes. Varying numbers of primary porcine chondrocytes (0.25, 2.5, 25 and 250 K) were seeded in 96 well-plates and cultured for 4 weeks. Cell proliferation, glycosaminoglycan (GAG) production and gene expression levels of Sox9, aggrecan, COL II and COL I were evaluated. The results showed that GAG content was high in the 0.25 and 25 K groups, gene expression of Sox9 was high in the 2.5, 25 and 250 K groups and expression of COL II was high in the 25 K group, whereas expression of COL I was low in the 0.25, 25 and 250 K groups. It is concluded that the seeding number and density of the 25 K (78 K cells/cm2) group achieved the optimal balance between functional phenotype of individual cells and the total ECM production for in vitro cultured chondrocytes.

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“…As a terminal marker, such an early expression seems questionable. Furthermore, Col 10 , being usually associated with the hypertrophic cartilage zone, was also detected in native cartilage (Bernstein et al ., ). Other studies concluded that ongoing Sox 9 expression, as detected in our cultures, might inhibit a hypertrophic development despite Col 10 expression (Mueller and Tuan, ; Mayer‐Wagner et al ., ).…”

Section: Discussionmentioning

confidence: 97%

Modulation of chondrogenic differentiation of human mesenchymal stem cells in jellyfish collagen scaffolds by cell density and culture medium

Pustlauk

Paul

Brueggemeier

et al. 2015

J Tissue Eng Regen Med

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Studies on tissue-engineering approaches for the regeneration of traumatized cartilage focus increasingly on multipotent human mesenchymal stem cells (hMSCs) as an alternative to autologous chondrocytes. The present study applied porous scaffolds made of collagen from the jellyfish Rhopilema esculentum for the in vitro chondrogenic differentiation of hMSCs. Culture conditions in those scaffolds differ from conditions in high-density pellet cultures, making a re-examination of these data necessary. We systematically investigated the influence of seeding density, basic culture media [Dulbecco's modified Eagle's medium (DMEM), α-minimum essential medium (α-MEM)] with varying glucose content and supplementation with fetal calf serum (FCS) or bovine serum albumin (BSA) on the chondrogenic differentiation of hMSCs. Gene expression analyses of selected markers for chondrogenic differentiation and hypertrophic development were conducted. Furthermore, the production of cartilage extracellular matrix (ECM) was analysed by quantification of sulphated glycosaminoglycan and collagen type II contents. The strongest upregulation of chondrogenic markers, along with the highest ECM deposition was observed in scaffolds seeded with 2.4 × 10 cells/cm after cultivation in high-glucose DMEM and 0.125% BSA. Lower seeding densities compared to high-density pellet cultures were sufficient to induce in vitro chondrogenic differentiation of hMSCs in collagen scaffolds, which reduces the amount of cells required for the seeding of scaffolds and thus the monolayer expansion period. Furthermore, examination of the impact of FCS and α-MEM on chondrogenic MSC differentiation is an important prerequisite for the development of an osteochondral medium for simultaneous osteogenic and chondrogenic differentiation in biphasic scaffolds for osteochondral tissue regeneration. Copyright © 2015 John Wiley & Sons, Ltd.

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Sox9 expression of alginate-encapsulated chondrocytes is stimulated by low cell density

Abstract: The authors regret the error.

Cited by 5 publications

References 0 publications

Effects of cell type and configuration on anabolic and catabolic activity in 3D co‐culture of mesenchymal stem cells and nucleus pulposus cells

Effects of cell type and configuration on anabolic and catabolic activity in 3D co‐culture of mesenchymal stem cells and nucleus pulposus cells

RELATIONSHIP BETWEEN CELL FUNCTION AND INITIAL CELL SEEDING DENSITY OF PRIMARY PORCINE CHONDROCYTES IN VITRO

Modulation of chondrogenic differentiation of human mesenchymal stem cells in jellyfish collagen scaffolds by cell density and culture medium

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