scholarly journals High-throughput, temporal and dose dependent, effect of vitamins and minerals on chondrogenesis

2019 ◽  
Author(s):  
James E. Dennis ◽  
Taylor Splawn ◽  
Thomas J. Kean
Keyword(s):  
High Throughput ◽  
Hyaline Cartilage ◽  
Culture Media ◽  
Fold Increase ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Collagen Expression ◽  
Culture Model ◽  
Engineered Tissue ◽  
Collagen Promoter

AbstractTissue engineered hyaline cartilage is plagued by poor mechanical properties largely due to inadequate type II collagen expression. Of note, commonly used defined chondrogenic media lack 14 vitamins and minerals, some of which are implicated in chondrogenesis. Type II collagen promoter-driven Gaussia luciferase was transfected into ATDC5 cells to create a chondrogenic cell with a secreted-reporter. The reporter cells were used in an aggregate-based chondrogenic culture model to develop a high-throughput analytic platform. This high-throughput platform was used to assess the effect of vitamins and minerals, alone and in combination with TGFβ1, on type II collagen expression. Significant combinatorial effects between vitamins, minerals and TGFβ1 in terms of type II collagen expression and metabolism were discovered. An ‘optimal’ continual supplement of copper and vitamin K in the presence of TGFβ1 gave a 2.5-fold increase in collagen expression over TGFβ1 supplemented media alone.SummaryCurrent defined chondrogenic culture media lack several vitamins and minerals. Type II collagen is the quintessential marker of articular hyaline cartilage, and is commonly deficient in engineered tissue. A type II collagen promoter driven secreted luciferase construct has been transduced into ATDC5 cells and used to assess vitamin and mineral effects on chondrogenesis in a high-throughput format.

Type VI collagen expression is upregulated in the early events of chondrocyte differentiation

Development ◽  
1993 ◽  
Vol 117 (1) ◽  
pp. 245-251
Author(s):  
R. Quarto ◽  
B. Dozin ◽  
P. Bonaldo ◽  
R. Cancedda ◽  
A. Colombatti
Keyword(s):  
Suspension Culture ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Type I ◽  
Type Ii ◽  
Type Vi Collagen ◽  
Full Spectrum ◽  
Collagen Expression ◽  
Hypertrophic Chondrocyte

Dedifferentiated chondrocytes cultured adherent to the substratum proliferate and synthesize large amounts of type I collagen but when transferred to suspension culture they decrease proliferation, resume the chondrogenic phenotype and the synthesis of type II collagen, and continue their maturation to hypertrophic chondrocyte (Castagnola et al., 1986, J. Cell Biol. 102, 2310–2317). In this report, we describe the developmentally regulated expression of type VI collagen in vitro in differentiating avian chondrocytes. Type VI collagen mRNA is barely detectable in dedifferentiated chondrocytes as long as the attachment to the substratum is maintained, but increases very rapidly upon passage of the cells into suspension culture reaching a peak after 48 hours and declining after 5–6 days of suspension culture. The first evidence of a rise in the mRNA steady-state levels is obtained already at 6 hours for the alpha 3(VI) chain. Immunoprecipitation of metabolically labeled cells with type VI collagen antibodies reveals that the early mRNA rise is paralleled by an increased secretion of type VI collagen in cell media. Induction of type VI collagen is not the consequence of trypsin treatment of dedifferentiated cells since exposure to the actin-disrupting drug cytochalasin or detachment of the cells by mechanical procedures has similar effects. In 13-day-old chicken embryo tibiae, where the full spectrum of the chondrogenic differentiation process is represented, expression of type VI collagen is restricted to the articular cartilage where chondrocytes developmental stage is comparable to stage I (high levels of type II collagen expression).(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Looping Mediated Interaction between the Promoter and 3′ UTR Regulates Type II Collagen Expression in Chondrocytes

PLoS ONE ◽  
2012 ◽  
Vol 7 (7) ◽  
pp. e40828 ◽  
Author(s):  
Arijita Jash ◽  
Kangsun Yun ◽  
Anupama Sahoo ◽  
Jae-Seon So ◽  
Sin-Hyeog Im
Keyword(s):  
Type Ii Collagen ◽  
Type Ii ◽  
Collagen Expression

scholarly journals Regulation of type II collagen expression by cyclin-dependent kinase 6, cyclin D1, and p21 in articular chondrocytes

IUBMB Life ◽  
2007 ◽  
Vol 59 (2) ◽  
pp. 90-98 ◽  
Author(s):  
Sang-Gu Hwang ◽  
Sung-Min Song ◽  
Jeong-Ran Kim ◽  
Chun-Shik Park ◽  
Woo-Keun Song ◽  
...  
Keyword(s):  
Cyclin D1 ◽  
Articular Chondrocytes ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Cyclin Dependent Kinase ◽  
Collagen Expression

scholarly journals Cellular heterogeneity in cultured human chondrocytes identified by antibodies specific for alpha 2(XI) collagen chains.

1989 ◽  
Vol 109 (3) ◽  
pp. 1363-1369 ◽  
Author(s):  
B Swoboda ◽  
R Holmdahl ◽  
H Stöss ◽  
K von der Mark
Keyword(s):  
Hyaline Cartilage ◽  
Type Ii Collagen ◽  
Cross Reactivity ◽  
Cellular Heterogeneity ◽  
Human Chondrocytes ◽  
Type I ◽  
Type Ii ◽  
Human Cartilage ◽  
The Matrix ◽  
Type Xi Collagen

Collagen type XI is a component of hyaline cartilage consisting of alpha 1(XI), alpha 2(XI), and alpha 3(XI) chains; with 5-10% of the total collagen content, it is a minor but significant component next to type II collagen, but its function and precise localization in cartilaginous tissues is still unclear. Owing to the homology of the alpha 3(XI) and alpha 1(II) collagen chains, attempts to prepare specific antibodies to native type XI collagen have been unsuccessful in the past. In this study, we report on the preparation and use for immunohistochemistry of a polyclonal antibody specific for alpha 2(XI) denatured collagen chains. The antibody was prepared by immunization with the isolated alpha 2(XI) chain and reacts neither with native type XI collagen nor type I, II, V, or IX by ELISA or immunoblotting, nor with alpha 1(XI) or alpha 3(XI), but with alpha 2(XI) chains. Using this antibody, it was possible to specifically localize alpha 2(XI) in cartilage by pretreating tissue sections with 6 M urea. In double immunofluorescence staining experiments, the distribution of alpha 2(XI) as indicative for type XI collagen in fetal bovine and human cartilage was compared with that of type II collagen, using a monoclonal antibody to alpha 1(II). Type XI collagen was found throughout the matrix of hyaline cartilage. However, owing to cross-reactivity of the monoclonal anti-alpha 1(II) with alpha 3(XI), both antibodies produced the same staining pattern. Cellular heterogeneity was, however, detected in monolayer cultures of human chondrocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

SOX9 expression does not correlate with type II collagen expression in adult articular chondrocytes

2003 ◽  
Vol 22 (4) ◽  
pp. 363-372 ◽  
Author(s):  
Thomas Aigner ◽  
Pia Margarethe Gebhard ◽  
Erik Schmid ◽  
Brigitte Bau ◽  
Vincent Harley ◽  
...  
Keyword(s):  
Articular Chondrocytes ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Sox9 Expression ◽  
Collagen Expression
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