Synthesis of chondrocytic keratan sulphate-containing proteoglycans by human chondrosarcoma cells in long-term cell culture.

Block, Joel A.; Inerot, Sven; Gitelis, Steven; Kimura, James H.

doi:10.2106/00004623-199173050-00002

Cited by 62 publications

(32 citation statements)

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“…In addition, monolayer 2-dimensional culture induces loss of the differentiated phenotype, as characterized by the upregulation of col1 and the downregulation of col2 [Benya and Shaffer, 1982;Zwicky and Baici, 2000]. Some previously created immortalized cell lines have been incapable of producing col2 [Takigawa et al, 1989;Block et al, 1991;Thenet et al, 1992], and therefore both the phenotype and morphology of A13/BACii were investigated and compared to freshly isolated bovine chondrocytes and the col2-producing cell line C-20/A4 [Goldring et al, 1994]. As 3-dimensional encapsulation into supports composed of col2, agarose or alginate enables the recapture of phenotype [Beekman et al, 1997;Nehrer et al, 1997;Benz et al, 2002;Miot et al, 2005], we investigated the changes in both the morphology and 'expression battery' of A13/BACii in response to HD low-melt agarose 3-dimensional culture as previously described [Schulze-Tanzil et al, 2002].…”

Section: Discussionmentioning

confidence: 99%

The Phenotypic Characterization of A13/BACii, a Novel Bovine Chondrocytic Cell Line with Differentiation Potential

Qusous

Kaneva

Can

et al. 2012

Cells Tissues Organs

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In cartilage research bovine articular cartilage is used as an alternative to human tissue. However, animal material is subject to availability and primary cultures undergo senescence, limiting their use. Here we report the immortalization of primary bovine chondrocytes, which could be used as a surrogate for freshly isolated chondrocytes. Chondrocytes were isolated from cartilage explants and immortalized using 1.0 µg/ml benzo[alpha]pyrene. For 3-dimensional culture, chondrocytes were resuspended in 0.5% low-melt agarose at high density (HD) and cultured for 24 h prior to determining changes in expression profile and morphology. A13/BACii chondrocytes acquired a ‘flat’ irregular morphology and a foetal-like cell volume (1,509.59 ± 182.04 µm³). The human cell line C-20/A4 showed a statistically similar volume and length to A13/BACii. Two-dimensional-cultured A13/BACii expressed elevated levels of type I collagen (col1), reduced levels of type II collagen (col2) compared to freshly isolated chondrocytes and an overall col2 to col1 expression ratio (col2:col1) of 0.11 ± 0.01. Upon 3-dimensional encapsulation, there was a significant rise in col2 expression in both A13/BACii and C-20/A4, suggesting a capacity for redifferentiation in both cell lines with a return of col2:col1 values of A13/BACii to values previously observed in primary chondrocytes. A13/BACii chondrocytes expressed aggrecan, matrix metalloproteinase (MMP)-3, MMP-9 and MMP-13, further supporting indications of the differentiated phenotype. Here we report the creation of a novel chondrocytic cell line and demonstrate its strong potential for redifferentiation upon HD 3-dimensional encapsulation, providing an alternative to conventional dedifferentiated cell lines and primary culture.

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Section: Discussionmentioning

confidence: 99%

The Phenotypic Characterization of A13/BACii, a Novel Bovine Chondrocytic Cell Line with Differentiation Potential

Qusous

Kaneva

Can

et al. 2012

Cells Tissues Organs

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“…Two lines of evidence suggest that regulation of GAG chain attachment may be important during tissue development. It has been reported that attachment of GAG chains changes during tissue growth, and the types and numbers of GAG chains vary during different growth stages (34). Furthermore, the GAG chains of versican have been reported to be involved in reducing cell adhesion (35).…”

Section: Figmentioning

confidence: 99%

Tandem Repeats Are Involved in G1 Domain Inhibition of Versican Expression and Secretion and the G3 Domain Enhances Glycosaminoglycan Modification and Product Secretion via the Complement-binding Protein-like Motif

Yang¹,

Cao²,

Kiani

et al. 2000

Journal of Biological Chemistry

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The large aggregating chondroitin sulfate proteoglycans, including aggrecan, versican (PG-M), neurocan, and brevican, are characterized by N-terminal and Cterminal globular (or selectin-like) domains known as the G1 and G3 domains, respectively. For this study, we generated a series of expression constructs containing various combinations of chicken versican/PG-M domains and a leading peptide of link protein in order to examine the roles of the G1 and G3 domains in versican function. In transfection studies, we observed that the presence of the G1 domain was sufficient to inhibit product secretion, while the G3 domain enhanced this process. We also demonstrated that the G1 domain inhibited the attachment of glycosaminoglycan chains to the core proteins, while the G3 domain enhanced this process. Further studies revealed that inhibition of secretion by G1 was mediated by its two tandem repeats, while G3's promotion of glycosaminoglycan chain attachment was apparently dependent on G3's complement-binding protein (CBP)-like motif. The modulatory effects of these two molecular domains may contribute to versican's biological activities.

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“…These characteristics are unique compared with the cell lines with cartilaginous phenotypes reported previously [4,10,11,24,45].…”

Section: Discussionmentioning

confidence: 84%

“…It is, therefore, important to establish a model of CS with a chondrosarcomatous phenotype for clinical investigation. Until now, there have been some reported cell lines of CS; however, those with chondrocytic phenotypes were limited [4,10,11,24,45]. In this study, new cell lines with chondrocytic phenotypes were established from a pelvic chondrosarcoma, and the morphological and molecular analyses were performed.…”

Section: Introductionmentioning

confidence: 99%

New cell lines with chondrocytic phenotypes from human chondrosarcoma

et al. 2004

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In the present study, we investigated chondrocytic characterization for newly established human chondrosarcoma cell lines. A chondrosarcoma cell line, HCS-TG, was established by the implantation of grade-2 human chondrosarcoma into athymic mice. Cloning of HCS-TG cells from passage 17 was performed. After cell cloning, two clonal-cell lines (HCS-TG C3 and E2) with good proliferative activities were obtained. These cell lines in monolayer culture retained a polygonal morphology. Their doubling times were 68 h and 45 h, respectively. mRNA expressions of type-I, -II, -X, and -XI collagens and aggrecan core protein were detected on reverse-transcription polymerase chain reaction. Protein expression of type-II collagen was confirmed in each cell line using Western blotting. However, there was no expression of type-I collagen. Moreover, gelatin zymography revealed that both cell lines produced extracellular matrices with matrix metalloproteinases 2 and 9. The parental HCS-TG cells had tumorigenicity in athymic mice; however, C3 and E2 were not tumorigenic. New clonal-cell lines HCS-TG C3 and E2 derived from human chondrosarcoma are morphologically chondrocytic in serial monolayer cultures and express chondrocytic phenotypes.

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Synthesis of chondrocytic keratan sulphate-containing proteoglycans by human chondrosarcoma cells in long-term cell culture.

Cited by 62 publications

References 0 publications

The Phenotypic Characterization of A13/BACii, a Novel Bovine Chondrocytic Cell Line with Differentiation Potential

The Phenotypic Characterization of A13/BACii, a Novel Bovine Chondrocytic Cell Line with Differentiation Potential

Tandem Repeats Are Involved in G1 Domain Inhibition of Versican Expression and Secretion and the G3 Domain Enhances Glycosaminoglycan Modification and Product Secretion via the Complement-binding Protein-like Motif

New cell lines with chondrocytic phenotypes from human chondrosarcoma

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