A quantitative method for analysis of radiolabelled proteoglycans synthesized by cultured human arterial smooth muscle cells.
1. Sulphate labelled proteoglycans (PG) synthesized by cultured human arterial smooth muscle cell have been quantified using an improved method based on a combination of specific enzymes and ethanol precipitation. 2. The present method gives quantitative data of PGs and subclasses allowing batchwise analysis of a large number of samples. 3. Approximately 81% +/- 1.7% (mean +/- SD, n = 6) of total PGs synthesized by human arterial smooth muscle cells accumulated in medium. 4. In cell layer and medium chondroitin sulphate proteoglycan constituted 65.0% +/- 0.3% and 75.8% +/- 0.7% (mean +/- SD, n = 3), respectively of sulphated PGs. 5. Heparan sulphate proteoglycan accounted for 26.8% +/- 0.6% in cell layer and 22.6% +/- 0.5% (mean +/- SD, n = 3) in medium of sulphated PGs.
Thøgersen
V B
VB
Research Laboratory of Biochemical Pathology, Aarhus Kommunehospital, University of Aarhus, Denmark.
Heickendorff
L
L
Ledet
T
T
D001011:Aorta; D002478:Cells, Cultured; D011232:Chemical Precipitation; D002847:Chromatography, Agarose; D002852:Chromatography, Ion Exchange; D000431:Ethanol; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism*; D013462:Sulfur Radioisotopes
D011509:Proteoglycans; D013462:Sulfur Radioisotopes; D000431:Ethanol