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Updated: Jul 17, 2026
Measuring Proliferation of Vascular Smooth Muscle Cells Using Click Chemistry
Published on: October 30, 2019
Iododeoxyuridine uptake in proliferating smooth muscle cells in vitro
Yonghua Xu1, Mandar R Jagtap, Tam Garland
1Department of Radiology, Indiana University Medical Center, Indianapolis, IN 47405, USA.
Purpose:
Iododeoxyuridine (IUdR) is a halogenated pyrimidine recognized as the thymidine substitute in DNA. When labeled with iodine 125, IUdR can be used as a carrier to incorporate the isotope into DNA and target the dividing cells. The purpose of this study was to assess the maximum uptake of IUdR by proliferating smooth muscle cells (SMCs) in vitro to determine the optimal concentration to be administered in an in vivo experiment. The long-term goal is to use radioactive IUdR to inhibit SMC proliferation and recurrent stenosis of arteries after balloon angioplasty in vivo.
Materials And Methods:
Porcine vascular SMCs were cultured in 5% fetal bovine serum medium and stimulated to proliferate by adding a medium containing 10% fetal bovine serum and insulin. IUdR was added to the proliferating SMCs at concentrations of 5, 10, 20, 30, and 40 micro mol/L on days 1, 3, 5, and 7 of incubation. One group of cells--the control group--did not receive IUdR. The SMCs were harvested and double-stained with an anti-IUdR antibody and propidium iodide, and fluorescence-activated cell scanning was performed to determine the ratio of IUdR-labeled cells to the total cell population for each IUdR concentration and at each time point. The data were measured three times at each time point. The doubling times, growth curve, and cell density of the proliferating SMCs were investigated by using the Coulter particle counter and digital microscopy.
Results:
The percentage of proliferating SMCs that showed IUdR uptake increased from 1 to 5 days incubation with all concentrations of IUdR; the incorporation rate reached a peak value at day 5 and then decreased by day 7. IUdR uptake on day 5 was higher with concentrations of 10 and 20 micro mol/L. When compared with that of the control group, the doubling times increased with an increase in IUdR concentration, whereas the proliferating cell number and density decreased significantly by days 5 (P < .05) and 7 (P < .01).
Conclusions:
IUdR uptake peaked on day 5, and the optimal concentration of IUdR for in vitro uptake in proliferating SMCs was 10-20 micro mol/L. IUdR inhibited the proliferation of the SMCs, and the inhibitory effect was related to the concentration.
