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Isolation og in vitro-kultur af knoglemarvsafledte makrofager til undersøgelse af NO-Redox-biologi
JoVE Journal
Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Isolation and In vitro Culture of Bone Marrow-Derived Macrophages for the Study of NO-Redox Biology

Isolation og in vitro-kultur af knoglemarvsafledte makrofager til undersøgelse af NO-Redox-biologi

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

Capitoli

  • 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

Traduzione automatica

Traduzione automatica

Denne protokol er blevet etableret for at dyrke tetrahydrobiopterin (BH4) - og inducerbar nitrogenoxidsyntase (iNOS) - mangelfulde primære murine makrofager til at studere NO-redox biologi. Undersøgelsen fokuserer på at reducere potentiel forurening af BH4 og andre artefakter, der findes i traditionelle isolations- og kulturmetoder, som kan forvirre eksperimentelle resultater og fortolkning af resultater.

Tags

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

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