ArticleAnnals of translational medicine2022
Bone marrow is the preferred source for isolation of rat neutrophils and the subsequent acquisition of neutrophil extracellular traps.
Article in Annals of translational medicine, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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4 citing papers in PubMed.
- Lactate-Induced Mitochondrial Calcium Uptake 3 Aggravates Myocardial Ischemia-Reperfusion Injury by Promoting Neutrophil Extracellular Trap Formation.Research (Washington, D.C.) · 2025Article
- Article
- Interaction between neutrophil extracellular traps and cardiomyocytes contributes to atrial fibrillation progression.Signal transduction and targeted therapy · 2023Article
- Abnormal adenosine metabolism of neutrophils inhibits airway inflammation and remodeling in asthma model induced by Aspergillus fumigatus.BMC pulmonary medicine · 2023Article
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6 authors.
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Abstract
Background: Neutrophil extracellular traps (NETs) are a network structure with DNA as skeleton scaffolding a wide range of antimicrobial proteins, which have been shown to contribute to the pathogenesis, persistence, and progression of many chronic inflammatory diseases. The method for isolation of human or mouse NETs has been well-established. However, in some diseases such as atrial fibrillation, the model can only be established in rats. While most related studies isolate rat neutrophils from the peripheral blood, which is insufficient for further acquisition of NETs. Despite the consumptiveness of rat peripheral blood neutrophil isolation, bone-marrow deprived neutrophil is rarely employed and has not been compared to peripheral neutrophil with its NETs secreting capability. Methods: In the current study, a different bone-marrow-oriented strategy was described and conducted. Based on centrifugal program settings, a one-step method and a two-step method was developed and compared. The purity of the isolated neutrophils was determined by Wright's staining and the viability was detected by flow cytometry. NETosis is the specialized cell death of neutrophils accompanied with NETs formation and its degree of rat neutrophils was analyzed by phase contrast microscopy, fluorescence microscopy, and Celigo whole view analysis. The Picogreen dsDNA Assay Kit was used to determine the concentration of NETs obtained from the NET-rich supernatants. The levels of secreted NETs in rat peripheral blood and bone marrow were compared. Results: Approximately 0.5×10 Conclusions: The bone marrow-oriented strategy for isolating rat neutrophils is accessible and repeatable. NETs formation capability is similar between rat bone-marrow and peripheral blood neutrophils.
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