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Biochemistry

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JoVE Journal Biochemistry
A Semi-Automated and Reproducible Biological-Based Method to Quantify Calcium Deposition In Vitro
 

A Semi-Automated and Reproducible Biological-Based Method to Quantify Calcium Deposition In Vitro

Article DOI: 10.3791/64029-v • 11:30 min • June 2nd, 2022
June 2nd, 2022 •
*1, *1, 1, 2, 1, 2, 1,3
1Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, 2Helmholtz Institute for Biomedical Engineering, Biointerface Group, RWTH Aachen University, 3Institute of Experimental Medicine and Systems Biology, RWTH Aachen University
* These authors contributed equally

Kapitel

Summary
Automatic Translation

Cardiovascular disease is the leading cause of death worldwide. Vascular calcification contributes substantially to the burden of cardiovascular morbidity and mortality. This protocol describes a simple method to quantify vascular smooth muscle cell-mediated calcium precipitation in vitro by fluorescent imaging.

Tags

Semi-automated Reproducible Biological-based Method Quantify Calcium Deposition In Vitro Calcification Propensity Patient Samples Human Vascular Smooth Muscle Cells Vascular Calcification Disorders New Therapies Fluorescent-based Detection Sensitivity Measure Samples Over Time Reducing Replicates Increasing Robustness Culture Human Vascular Smooth Muscle Cells Uncoated Cell Culture Flasks 37 Degrees Celsius 5% Carbon Dioxide Split Cells Wash With PBS Trypsin Reaction Serum-containing Medium Centrifuge Cells Resuspend Pellet Maintenance Medium Count And Seed Cells 48-well Plate Calcification Experiment Overnight Incubation Wash With PBS Aspirate Remaining PBS Add Calcification Medium
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