Preliminary experience with cell culture of human articular chondrocytes

Srivastava, V. M. L.; Malemud, Charles J.; Hough, Aubrey J.; Bland, John H.; Sokoloff, Leon

doi:10.1002/art.1780170209

Cited by 42 publications

(11 citation statements)

References 13 publications

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“…These observations reemphasize the necessity for rigid control of experimental design when comparing parameters for different cell types under varying culture conditions. Ascorbic acid has been shown to stimulate sulfated proteoglycan biosynthesis in chick embryo fibroblasts (46), have no effect in rabbit chondrocytes (44), and to cause decreased sulfate incorporation in human articular cartilage chondrocytes (47).…”

Section: Resultsmentioning

confidence: 99%

Effect of ascorbic acid on arylsulfatase activities and sulfated proteoglycan metabolism in chondrocyte cultures.

Schwartz¹,

Adamy²

1977

J. Clin. Invest.

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A B S T R A C T A correlation between increased arylsulfatase activities and decreased sulfated proteoglycan content in human osteoarthritic articular cartilage suggested a possible interrelationship between these parameters. Since we had previously shown that ascorbate caused a decrease in levels of arylsulfatase A and B activities in normal chondrocyte cultures, the validity of the above relationship was examined by measuring the effect of vitamin C on the biosynthesis and distribution of 35S-labeled proteoglyeans and arylsulfatase A and B activities in cell extracts of chondrocytes derived from normal and osteoarthritic tissue.Arylsulfatase A and B activities were found to be reduced in the presence of ascorbic acid in all normal and osteoarthritic cell lines examined when measured 3, 6, 10, and 13 days after the introduction of the vitamin in the culture medium. Acid phosphatase activity, on the other hand, was found to be elevated in the presence of ascorbate.The inhibitory effect by ascorbic acid on arylsulfatase activities could be reversed by withdrawing the vitamin from the nutrient medium. Addition of EDTA to the cell extracts before assay also reversed the inhibition.Sulfated proteoglycan biosynthesis as reflected in 35S-sulfate uptake per milligram of DNA was significantly increased in the presence of ascorbic acid. The distribution of the newly synthesized molecules between the cell layer and medium fractions was altered. In the presence of ascorbate, more deposition into the cell layer of newly synthesized macromolecules occurred.These data suggest an inverse relationship between arylsulfatase activities and the stability of the newly synthesized sulfated proteoglycans in the extracellular matrix.

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Section: Resultsmentioning

confidence: 99%

Effect of ascorbic acid on arylsulfatase activities and sulfated proteoglycan metabolism in chondrocyte cultures.

Schwartz¹,

Adamy²

1977

J. Clin. Invest.

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“…We chose to use 10% FBS as our standard because this concentration is most often used for culturing various types of chondrocytes (1, 5,11,19,31,32,35,38,40), including rabbit meniscal fibrochondrocytes (37). The final formula developed was quite different in many respects from formulations found to be useful in supporting the proliferation of other chondrocyte types.…”

Section: Discussionmentioning

confidence: 99%

Serum‐free culture of rabbit meniscal fibrochondrocytes: Proliferative response

Webber

Zitaglio

Hough

1988

Journal Orthopaedic Research

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We have formulated a serum-free medium capable of supporting DNA synthesis in rabbit meniscal fibrochondrocytes at a level equivalent to 10% fetal bovine serum (FBS). The medium consists of a 1:1 mixture of Dulbecco's Modified Eagle's Medium and Ham's F-12 medium supplemented with transferrin (1 microgram/ml), selenium (1 pg/ml), trace metal mix (1:100), dexamethasone (100 ng/ml), insulin-like growth factors I and II (50 ng/ml each), pituitary fibroblast growth factor (100 ng/ml), and lactalbumin hydrolysate (2 micrograms/ml). Endothelial cell growth supplement could be substituted for lactalbumin hydrolysate to obtain similar results. Ventrex PC-1, a commercially available, low-protein, serum-free medium, was found to support proliferation of fibrochondrocytes but not as well as 10% FBS or our medium formulation. Lipid supplements, which are known to support the serum-free growth of hyaline chondrocytes, were found to be either of no value or antagonistic for the culture of fibrochondrocytes. Likewise, vitamin E alone, progesterone, putrescine, and hydrocortisone were also without benefit in our culture system. The cells had a more chondrocytic morphology when grown in defined medium as opposed to 10% FBS. The results of this study should now make it possible to identify and quantitate those factors necessary to affect meniscal repair by utilizing further techniques in vitro.

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“…However, the previous reports are conflicting. In cultured human articular chondrocytes, ascorbic acid (50 pg/ml) has been shown to produce a 2-fold decrease in glycosaminoglycan (GAG) secretion [8] whereas it increases this latter by 70% when used at 70 fig/ml [9]. Addition of ascorbic acid (40pg/ml) to cultures of rabbit artitular chondrocytes obtamed by enzymatic dissoclatron does not change the 35S incorporation m proteoglycans [lO,ll].…”

Section: Introductionmentioning

confidence: 99%

Effect of asorbic acid on secreted proteoglycans from rabbit articular chondrocytes

et al. 1985

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Addrtron of ascorbrc acid (25, 50 100 pg/ml) to cultures of rabbrt articular chondrocytes did not change the total amount of proteoglycans produced However, It mduced an increased retention of these macromolecules m the pencellular fraction The size of the proteoglycan subumts and the length of glycosammoglycan chains, released m the medmm, were not moddied on exposure to ascorbic acid (25 pg/ml) On the other hand, the rate of non-sulfated chondroltm was increased 2 5-fold, whereas chondroltm4sulfate was depressed 1 5-fold ProteoglycanGlycosammoglycan

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Preliminary experience with cell culture of human articular chondrocytes

Cited by 42 publications

References 13 publications

Effect of ascorbic acid on arylsulfatase activities and sulfated proteoglycan metabolism in chondrocyte cultures.

Effect of ascorbic acid on arylsulfatase activities and sulfated proteoglycan metabolism in chondrocyte cultures.

Serum‐free culture of rabbit meniscal fibrochondrocytes: Proliferative response

Effect of asorbic acid on secreted proteoglycans from rabbit articular chondrocytes

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