Three-Dimensional Microenvironments Retain Chondrocyte Phenotypes During Proliferation Culture

Takahashi, Tatsuji; Ogasawara, Toru; Asawa, Yukiyo; Mori, Yoshiyuki; Uchinuma, Eiju; Takato, Tsuyoshi; Hoshi, Kazuto

doi:10.1089/ten.2006.0322

Cited by 96 publications

(70 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Pellet culture is a 3D cell culture system which has been demonstrated to be a good model for studies involving processes such as tissue induction and cellular differentiation [Johnstone et al, 1998;Le Visage et al, 2006;Takahashi et al, 2007]. This culture system mimics several different phases of tissue development and results in a differentiated, matrix-producing cell type.…”

Section: Discussionmentioning

confidence: 99%

Human Disk Cells from Degenerated Disks and Mesenchymal Stem Cells in Co-Culture Result in Increased Matrix Production

Svanvik¹,

Henriksson

Karlsson

et al. 2009

Cells Tissues Organs

View full text Add to dashboard Cite

Transplantation of mesenchymal stem cells (MSCs) has been suggested for disk degeneration, which is characterized by dysfunctional cells and low proteoglycan production. The aim of this study was to examine the effects of a 3D co-culture system using human disk cells (DCs) and MSCs on collagen and proteoglycan production. DCs and MSCs were expanded in monolayer and grown in pellet cultures for 7, 14 and 28 days and analyzed for hydroxyproline (HP), reflecting total collagen production, and glycosaminoglycan (GAG) accumulation. DCs and MSCs co-cultured at different ratios (25/75, 50/50 and 75%/25%) were examined for GAG accumulation. Collagen type II expression was analyzed immunohistochemically. In a second series, conditioned media were added to pellet cultures of degenerated DCs or MSCs. DCs from degenerated disks and MSCs demonstrated lower total collagen production than non-degenerated DC pellets. GAG production was comparable in DCs and MSCs, except in the youngest donor, with MSC producing about 10 times higher GAG/DNA. Co-cultures resulted in approximately 1.5 times higher GAG/DNA production than DCs. Increased collagen type II expression was seen in co-cultures compared to DC or MSC culture alone, except in the case with highly active MSCs. No positive effect of conditioned media was seen. In conclusion, co-culture of MSCs with degenerated DCs increased proteoglycan and collagen-type ceII production, indicating that in future clinical therapy MSCs can be transplanted without pre-differentiation in vitro. The lack of effect of conditioned media suggests that the positive effect of co-culture on matrix production is not due to soluble factors.

show abstract

Section: Discussionmentioning

confidence: 99%

Human Disk Cells from Degenerated Disks and Mesenchymal Stem Cells in Co-Culture Result in Increased Matrix Production

Svanvik¹,

Henriksson

Karlsson

et al. 2009

Cells Tissues Organs

View full text Add to dashboard Cite

show abstract

“…The COL2 evaluated in this study is a specific cartilage component. COL2 forms fibrils that permit cartilage to entrap proteoglycan aggregates and im- the surgical invasiveness of the biopsy and the physical strain on patients (22,29,31,32). Minimal invasiveness is one of the major goals of biomedical techniques, and our method may increase the advantages of this approach (33).…”

Section: Resultsmentioning

confidence: 99%

“…The chondrocytes were isolated by digestion with 0.3% collagenase over 24 h (Wako Pure Chemical Industries, Osaka, Japan). The primary auricular chondrocytes were seeded in 35-mm collagen type I-coated plastic culture dishes (n = 36) at a density of 5 × 10 3 cells/cm 2 and cultured in 2 mL of Dulbecco's modified Eagle's medium/F12 (DMEM/F12) containing 5% human serum supplemented with fibroblast growth factor-2 (100 ng/mL) and insulin (5 μg/mL) as previously described (10,22). We divided the 36 dishes into 3 groups (n = 12 per group).…”

Section: Methodsmentioning

confidence: 99%

Application of floating cells for improved harvest in human chondrocyte culture

Yonenaga¹,

Nishizawa²,

Fujihara³

et al. 2012

Biomed. Res.

Self Cite

View full text Add to dashboard Cite

Cell culture medium, which must be discarded during medium change, may contain many cells that do not attach to culture plates. In the present study, we focused on these floating cells and attempted to determine their usefulness for cartilage regeneration. We counted the number of floating cells discarded during medium change and compared the proliferation and differentiation between floating cells and their adherent counterparts. Chondrocyte monolayer culture at a density of 5 × 10 3 cells/cm 2 produced viable floating cells at a rate of 2.7-3.2 × 10 3 cells/cm 2 per primary culture. When only the floating cells from one dish were harvested and replated in another dish, the number of cells was 2. ). The floating and adherent cells showed similar proliferation and differentiation properties. The recovery of floating cells from the culture medium could provide an approximately 1.5-fold increase in cell number over conventional monolayer culture. Thus, the collection of floating cells may be regarded as a simple, easy, and reliable method to increase the cell harvest for chondrocytes.

show abstract

“…Encapsulation of dedifferentiated chondrocytes in alginate beads or the collagen gel is a common method in redifferentiating cells by promoting the synthesis of cartilage matrix proteins. 9,10 Chondrocyte culture in 3D, however, leads to a low rate of cell division. In addition, there are many reports that a dynamic culture environment is also helpful in maintaining the chondrocyte phenotype.…”

Section: Introductionmentioning

confidence: 99%

Induction of Re-Differentiation of Passaged Rat Chondrocytes Using a Naturally Obtained Extracellular Matrix Microenvironment

Hwa

Hee

Park

et al. 2013

Tissue Engineering Part A

View full text Add to dashboard Cite

Dedifferentiated human chondrocytes severely limit successful hyaline cartilage repair in clinical practice. The primary interest of this study is to evaluate the naturally obtained cell-derived matrix (CDM) as a physical microenvironment for chondrocyte re-differentiation. Once different cell types were cultured for 6 days and decellularized using detergents and enzymes, the fibroblast-derived matrix (FDM), preosteoblast-derived matrix (PDM), and chondrocyte-derived matrix (CHDM) were obtained. From scanning electron microscope observation, each CDM was found to resemble a fibrous mesh with self-assembled fibrils. Both the FDM and PDM showed a more compact matrix structure compared to the CHDM. For compositional analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis displayed numerous matrix proteins, which were quite different from each CDM in quantity and type. Specific matrix components, such as fibronectin, type I collagen (Col I), and laminin, were detected using immunofluorescent staining. In addition, the water contact angle suggests that the FDM is more hydrophilic than the PDM or CHDM. The proliferation of rat primary chondrocytes growing on CDMs was better than those growing on a plastic coverslip (control) or gelatin. Meanwhile, synthesis of glycosaminoglycan (GAG) was more effective for passaged chondrocytes (P4) cultivated on CDMs, and the difference was significant compared to cells grown on the control or on gelatin. As for the gene expression of cartilage-specific markers, CDMs exhibited good chondrocyte re-differentiation with time: the dedifferentiating marker, Col I was restrained, whereas the ratio between Col II and Col I, and between aggrecan and Col I, as an indicator of re-differentiation, was greatly improved. In addition, immunofluorescence of Col II showed a very positive signal in chondrocytes cultivated for 2 weeks on the CDMs. In an additional study, when threedimensional cell pellets made from either plate-grown or matrix-grown dedifferentiated chondrocytes (P5) were cultured for 4 weeks, the results of Safranin-O staining, immunohistochemistry of Col II, and total GAG assay suggested that matrix-grown cells were significantly better in the induction of chondrocyte re-differentiation, than those grown on the plate. This work suggests that the naturally occurring matrix, CDM, can provide a favorable surface texture for cell attachment, proliferation, and more importantly, a chondroinductive microenvironment for the re-differentiation of dedifferentiated chondrocytes.

show abstract

Three-Dimensional Microenvironments Retain Chondrocyte Phenotypes During Proliferation Culture

Cited by 96 publications

References 29 publications

Human Disk Cells from Degenerated Disks and Mesenchymal Stem Cells in Co-Culture Result in Increased Matrix Production

Human Disk Cells from Degenerated Disks and Mesenchymal Stem Cells in Co-Culture Result in Increased Matrix Production

Application of floating cells for improved harvest in human chondrocyte culture

Induction of Re-Differentiation of Passaged Rat Chondrocytes Using a Naturally Obtained Extracellular Matrix Microenvironment

Contact Info

Product

Resources

About