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Isolation and In vitro Culture of Bone Marrow-Derived Macrophages for the Study of NO-Redox Biology
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Isolation and In vitro Culture of Bone Marrow-Derived Macrophages for the Study of NO-Redox Biology

Isolation and In vitro Culture of Bone Marrow-Derived Macrophages for the Study of NO-Redox Biology

DOI:
10.3791/62834-v

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06:34 min

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May 31, 2022

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, , , , , , , ,
1BHF Centre of Research Excellence, Division of Cardiovascular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital,University of Oxford, 2Wellcome Centre for Human Genetics,University of Oxford

Chapters

  • 00:04Introduction
  • 00:53Isolation of Bone Marrow Cells
  • 02:14Selection, Differentiation, and Stimulation of BMDMs
  • 04:24Results: Comparison of Nitrite and Tetrahydrobiopterin Levels in Media Supplemented with LCM or Recombinant GM-CSF and M-CSF
  • 05:54Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol has been established to culture tetrahydrobiopterin (BH4)- and inducible nitric oxide synthase (iNOS)-deficient primary murine macrophages to study NO-redox biology. The study focuses on reducing potential contamination of BH4 and other artifacts found in traditional isolation and culture methods which may confound experimental outcomes and interpretation of results.

Tags

Keywords: Bone Marrow-derived MacrophagesINOSBH4Nitric OxideRedox BiologyIn Vitro CultureCell IsolationCell DisaggregationCell FreezingDMEM/F-12 Media

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