Cell-based semiquantitative assay for sulfated glycosaminoglycans facilitating the identification of chondrogenesis

Anal Biochem. 2015 Oct 1:486:41-3. doi: 10.1016/j.ab.2015.06.005. Epub 2015 Jun 9.

Abstract

Glycosaminoglycans (GAGs), in particular chondroitin sulfate, are an accepted marker of chondrogenic cells. In this study, a cell-based sulfated GAG assay for identifying the chondrogenesis of mesenchymal stem cells was developed. Based on fluorescent staining using safranin O and 4',6-diamidino-2-phenylindole (DAPI), this method was highly sensitive. The results were both qualitative and quantitative. The method is suitable for identifying the chondrogenic process and also for screening compounds. The method may be helpful for discovering novel bioactive compounds for cartilage regeneration.

Keywords: Chondrogenesis; DAPI; Glycosaminoglycan; Safranin O; Screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Child
  • Chondrogenesis / drug effects*
  • Chondroitin Sulfates / chemistry
  • Chondroitin Sulfates / metabolism*
  • DNA / chemistry
  • DNA / metabolism
  • Drug Evaluation, Preclinical / methods*
  • Humans
  • Indoles / chemistry
  • Limit of Detection
  • Mesenchymal Stem Cells / drug effects*
  • Mesenchymal Stem Cells / metabolism*
  • Phenazines / chemistry

Substances

  • Indoles
  • Phenazines
  • DAPI
  • Chondroitin Sulfates
  • DNA
  • safranine T