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In vitro Quantitative Imaging Assay for Phagocytosis of Dead Neuroblastoma Cells by iPSC-Macrophages
JoVE Journal
Neuroscienze
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JoVE Journal Neuroscienze
In vitro Quantitative Imaging Assay for Phagocytosis of Dead Neuroblastoma Cells by iPSC-Macrophages

In vitro Quantitative Imaging Assay for Phagocytosis of Dead Neuroblastoma Cells by iPSC-Macrophages

DOI:
10.3791/62217-v

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09:48 min

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February 14, 2021

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, , , ,
1James Martin Stem Cell Facility, Sir William Dunn School of Pathology,University of Oxford, 2Alzheimer’s Research UK Oxford Drug Discovery Institute, Nuffield Department of Medicine Research Building,University of Oxford, 3UK Dementia Research Institute,Cardiff University

Capitoli

  • 00:04Introduction
  • 00:41Preparation of Dead SH-SY5Ys
  • 02:03Labeling of Dead SH-SY5Ys with pH-Sensitive Red Fluorescent Dye
  • 03:18Staining of iPSC-Macrophages
  • 04:50Fixed-Cell High-Content Imaging
  • 07:28Results: Imaging, Optimization and Validation of Phagocytosis Assay
  • 09:16Conclusion

Summary

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Neurodegenerative diseases are associated with dysregulated microglia functions. This article outlines an in vitro assay of phagocytosis of neuroblastoma cells by iPSC-macrophages. Quantitative microscopy readouts are described for both live-cell time-lapse imaging and fixed-cell high-content imaging.

Tags

In VitroQuantitative Imaging AssayPhagocytosisDead Neuroblastoma CellsIPSC-macrophagesNeurodegenerative DiseaseSH-SY5Y CellsCell DissociationFixationPH-sensitive DyeFluorescent DyeMacrophage MediaHoechst 33342

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