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Chapters
- 00:00Title
- 00:57Introduction
- 01:58Preparing Anti-PECAM-1 Antibody-Conjugated Magnetic Beads
- 03:36Isolating Mouse Pulmonary Endothelial Cells from Neonatal Mice
- 06:56Sorting Mouse Lung Endothelial Cells with Anti-ICAM-2 Dynabeads
- 09:13Evaluation of Pulmonary Endothelial Cells from Neonatal Mice
- 09:57Conclusion
Here, we describe a protocol for isolation and culture of murine pulmonary endothelial cells. This method comprises mechanic and enzymatic lung tissue dissociation as well as a 2-step purification process using anti-PECAM-1 and anti-ICAM-2 antibodies conjugated to magnetic beads, which produces a pure endothelial cell population of mostly microvascular origin.
Tags
IsolationCulturePulmonary Endothelial CellsNeonatal MiceResearch ModelVascular BiologyHuman Umbilical Vein Endothelial Cells (HUVECs)Microvascular Endothelial Cells (ECs)AngiogenesisPermeabilityKnockout MiceProtein Function AnalysisIsolation ProtocolCulturing ProtocolMouse Lung Endothelial Cells (MLECs)Collagenase/dispase (C/D) SolutionSingle-cell SuspensionPositive SelectionAnti-PECAM-1 AntibodyDynabeadsMagnetic Particle Concentrator (MPC)
