Home › Analytical Chem › Kinetic Analysis of Vasculogenesis Quantifies Dynamics of Vasculogenesis and Angiogenesis In Vitro
Analytical Chem JoVE (Open Access) Citable · DOI

Kinetic Analysis of Vasculogenesis Quantifies Dynamics of Vasculogenesis and Angiogenesis In Vitro

DOI: 10.3791/57044-v
What you'll learn
  • ✓ Set up time-lapse phase contrast microscopy for vasculogenesis imaging
  • ✓ Apply Kinetic Analysis of Vasculogenesis software to quantify vascular dynamics
  • ✓ Measure and interpret kinetic parameters of vessel formation in vitro
  • ✓ Assess vasculogenic potential across different cell types or conditions
Protocol

Here, we present a protocol for time-lapse imaging and analysis of vasculogenesis in vitro using phase contrast microscopy coupled with the open source software, Kinetic Analysis of Vasculogenesis. This protocol can be applied to quantitatively assess the vasculogenic potential of numerous cell types or experimental conditions that model vascular disease.

Difficulty
intermediate
Total time
~3-5 days (including 24-48 hr culture period for vasculogenesis)
Model organism
Endothelial Colony Forming Cells (ECFCs)
Biosafety
BSL-1

Steps

1
Prepare microscope and plate ECFC matrix

Configure phase contrast microscopy system and prepare culture dishes with endothelial colony forming cells in appropriate matrix for vasculogenesis assay.

▶ 00:59
2
Acquire images and process with KAV software

Perform time-lapse image acquisition using phase contrast microscopy and process resulting image sequences using the Kinetic Analysis of Vasculogenesis open-source software.

▶ 04:46
3
Analyze kinetic parameters from vasculogenesis data

Review representative results and interpret kinetic analysis outputs to quantify vessel formation dynamics and compare vasculogenic potential across experimental conditions.

▶ 08:12
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