ArticleCalcified tissue international1995
A rapid and ultrasensitive method for measurement of DNA, calcium and protein content, and alkaline phosphatase activity of chondrocyte cultures.
Article in Calcified tissue international, 1995. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 48 citations in OpenAlex.
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- High-Frequency Acceleration: Therapeutic Tool to Preserve Bone following Tooth Extractions.Journal of dental research · 2016Article
- Investigation of chondrocyte hypertrophy and cartilage calcification in a full-depth articular cartilage explants model.Rheumatology international · 2013Article
- Intrinsically superparamagnetic Fe-hydroxyapatite nanoparticles positively influence osteoblast-like cell behaviour.Journal of nanobiotechnology · 2012Article
- Clonal growth of dermal papilla cells in hydrogels reveals intrinsic differences between Sox2-positive and -negative cells in vitro and in vivo.The Journal of investigative dermatology · 2012Article
- Article
- F-spondin regulates chondrocyte terminal differentiation and endochondral bone formation.Journal of orthopaedic research : official publication of the Orthopaedic Research Society · 2010Article
- An improved collagen scaffold for skeletal regeneration.Journal of biomedical materials research. Part A · 2010Article
- Interleukin-1beta and tumor necrosis factor alpha inhibit chondrogenesis by human mesenchymal stem cells through NF-kappaB-dependent pathways.Arthritis and rheumatism · 2009Article
- Engineering endochondral bone: in vitro studies.Tissue engineering. Part A · 2009Article
- Cell culture systems for studies of bone and tooth mineralization.Chemical reviews · 2008Review
- Partial reversal of transformed fusiform phenotype by overexpression of calreticulin.Cellular & molecular biology letters · 2007Article
- Retinoic acid modulation of glutathione and cysteine metabolism in chondrocytes.The Biochemical journal · 1996Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Most investigators are cognizant of the problems inherent in counting cells embedded in a complex and abundant extracellular matrix. To overcome these obstacles, we developed a new method of isolating nucleic acids from chondrocytes which facilitates measurement of cell number by DNA analysis. Chondrocytes were isolated from chick embryo sterna and grown continuously without subculturing for 2-3 weeks in monolayer. The cells were treated with triton X-100 and the nucleic acid content of the extract was determined by measuring DNA fluorescence in the presence of Hoechst dye 33258. To minimize background fluorescence due to the triton, we precipitated the DNA with alcohol and then solubilized the nucleic acids in EDTA. This simple procedure removed the detergent and substantially increased the sensitivity of the method. Thus, we could measure with high precision and high recovery, the DNA content of cultures of 10,000-50,000 cells. In a single well containing 0.5-1.0 million cells, sufficient material remained for subsequent measurements of alkaline phosphatase activity and protein and calcium content. As the mineral present in the triton-treated samples was soluble in EDTA, we experienced no problems in measuring the calcium content of the culture. In addition, as triton X-100 is a nonionic detergent, we were able to measure cell and matrix proteins; moreover, the presence of the triton maintained the catalytic state of alkaline phosphatase. We conclude that this procedure provides a simple and rapid approach to measuring major indicators of chondrocyte maturation and function.
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