Fluorometric assay of DNA in cartilage explants using Hoechst 33258
- PMID: 2464289
- DOI: 10.1016/0003-2697(88)90532-5
Fluorometric assay of DNA in cartilage explants using Hoechst 33258
Abstract
A simple two-step fluorometric assay of DNA in cartilage explants, utilizing the bisbenzimidazole dye Hoechst 33258, is described. Cartilage explants were prepared for assay by digestion with papain. Aliquots of the digest were mixed with dye solution, and the fluorescence emission measured. The enhancement in fluorescence of dye was specific for DNA, as demonstrated by 97% sensitivity to DNase and resistance to RNase. In addition, little or no interference was caused by non-DNA tissue components, since DNA caused an equal enhancement in fluorescence independent of the presence of papain-digested cartilage. By performing the assay on isolated chondrocytes, the cellular content of DNA was computed to be 7.7 pg per chondrocyte. The assay was stable for at least 2 h and sensitive to as little as 6 ng of DNA or equivalently less than 1000 cells. This procedure offers advantages over other established DNA assays of cartilage and may be especially useful in metabolic studies of cartilage explants.
Similar articles
-
Highly sensitive nuclease assays based on chemically modified DNA or RNA.Sensors (Basel). 2014 Jul 11;14(7):12437-50. doi: 10.3390/s140712437. Sensors (Basel). 2014. PMID: 25019631 Free PMC article. Review.
-
Biochemical quantification of DNA in human articular and septal cartilage using PicoGreen and Hoechst 33258.Osteoarthritis Cartilage. 2002 Jul;10(7):580-7. doi: 10.1053/joca.2002.0794. Osteoarthritis Cartilage. 2002. PMID: 12127839
-
Fluorometric quantification of DNA in cells and tissue.Anal Biochem. 1983 Jun;131(2):538-47. doi: 10.1016/0003-2697(83)90212-9. Anal Biochem. 1983. PMID: 6193739
-
Fluorophotometric quantitation of DNA in articular cartilage utilizing Hoechst 33258.Anal Biochem. 1989 Jan;176(1):128-31. doi: 10.1016/0003-2697(89)90282-0. Anal Biochem. 1989. PMID: 2469359
-
Fluorimetric determination of DNA in papain digests of cartilage, using ethidium bromide.Connect Tissue Res. 1979;6(4):215-21. doi: 10.3109/03008207909152323. Connect Tissue Res. 1979. PMID: 157845
Cited by
-
Enhanced hyaline cartilage formation and continuous osteochondral regeneration via 3D-Printed heterogeneous hydrogel with multi-crosslinking inks.Mater Today Bio. 2024 May 3;26:101080. doi: 10.1016/j.mtbio.2024.101080. eCollection 2024 Jun. Mater Today Bio. 2024. PMID: 38757056 Free PMC article.
-
Unlocking Potential: Low Bovine Serum Albumin Enhances the Chondrogenicity of Human Adipose-Derived Stromal Cells in Pellet Cultures.Biomolecules. 2024 Mar 28;14(4):413. doi: 10.3390/biom14040413. Biomolecules. 2024. PMID: 38672430 Free PMC article.
-
A synthetic, closed-looped gene circuit for the autonomous regulation of RUNX2 activity during chondrogenesis.FASEB J. 2024 Feb 29;38(4):e23484. doi: 10.1096/fj.202300348RR. FASEB J. 2024. PMID: 38407380
-
Type II collagen scaffolds repair critical-sized osteochondral defects under induced conditions of osteoarthritis in rat knee joints via inhibiting TGF-β-Smad1/5/8 signaling pathway.Bioact Mater. 2024 Feb 16;35:416-428. doi: 10.1016/j.bioactmat.2024.02.008. eCollection 2024 May. Bioact Mater. 2024. PMID: 38384986 Free PMC article.
-
The Phenotype of Mesenchymal Stromal Cell and Articular Chondrocyte Cocultures on Highly Porous Bilayer Poly-L-Lactic Acid Scaffolds Produced by Thermally Induced Phase Separation and Supplemented with Hydroxyapatite.Polymers (Basel). 2024 Jan 25;16(3):331. doi: 10.3390/polym16030331. Polymers (Basel). 2024. PMID: 38337220 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources