2014
DOI: 10.3791/52347
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Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes

Abstract: As a subtype of leukocytes and progenitors of macrophages, monocytes are involved in many important processes of organisms and are often the subject of various fields in biomedical science. The method described below is a simple and effective way to isolate murine monocytes from heterogeneous bone marrow.Bone marrow from the femur and tibia of Balb/c mice is harvested by flushing with phosphate buffered saline (PBS). Cell suspension is supplemented with macrophage-colony stimulating factor (M-CSF) and cultured… Show more

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Cited by 25 publications
(24 citation statements)
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“…As further evidence for the translatability of the approach, we also demonstrated tumor detection with primary BMDMs. We first generated monocytes from bone marrow and confirmed their phenotype by intermediate expression of the maturation marker F4/80 15 (Figure 4g). The pARG1-Gluc construct was introduced by electroporation with efficiencies greater than 80% (Supplementary Figure S1b) and the resulting sensor was activated ~10fold by both "low" and "high" TCM ( Figure 4h).…”
Section: An Engineered Macrophage Sensor Can Detect Sub 50 MM 3 Tumorsmentioning
confidence: 91%
See 1 more Smart Citation
“…As further evidence for the translatability of the approach, we also demonstrated tumor detection with primary BMDMs. We first generated monocytes from bone marrow and confirmed their phenotype by intermediate expression of the maturation marker F4/80 15 (Figure 4g). The pARG1-Gluc construct was introduced by electroporation with efficiencies greater than 80% (Supplementary Figure S1b) and the resulting sensor was activated ~10fold by both "low" and "high" TCM ( Figure 4h).…”
Section: An Engineered Macrophage Sensor Can Detect Sub 50 MM 3 Tumorsmentioning
confidence: 91%
“…Cells were maintained in a humidified, 5% CO 2 incubator at 37 °C for 5 days prior to harvesting with a cell lifter for downstream use. To make BMD monocytes 15 for all in vivo studies, cells were instead plated at a density of 6 × 10 6 cells/well in 6-well Corning® Costar® Ultra-Low Attachment Plates (Corning, Corning, NY) in 6 mL of media supplemented with 20 ng/mL M-CSF. After 5 days, adherent cells (macrophages) were discarded and the non-adhered cell were collected.…”
Section: Bone Marrow-derived Macrophage/monocyte (Bmdm) Preparation Amentioning
confidence: 99%
“…Macrophages were collected via mechanical dissociation with cell scrapers (VWR, Leuven, Belgium, #734-2602) after washing in 10 mM EDTA (Gibco, #15575-038) in DPBS and adding complete medium. Monocyte-derived cells intended to maintain a more immature monocyte status—hereafter referred to as ‘monocytes’—were cultured for only 5 days in 6-well ultra-low attachment plates (Sigma-Aldrich, Overijse, Belgium, #CLS3471-24EA) to prevent adherence-induced differentiation [18] . Culture medium was refreshed 3-4 days post-seeding.…”
Section: Methodsmentioning
confidence: 99%
“…NOTE: For the isolation of monocytes, please see our previous published video on JoVE for instructions: "Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes" by Wagner et al 4 NOTE: When working with the cells all steps must be sterile to avoid contamination.…”
Section: Cell Preparationmentioning
confidence: 99%
“…Monocytes are an important cell type associated with collateralization. Our group has focused on researching the effects of monocytes in areas of inflammation 4,5 , in particular using the hindlimb ischemia model to induce ischemia and subsequent inflammation 6 . Monocytes home to areas of inflammation and cause complex systemic responses that lead to the development of collateralization 7 .…”
Section: Introductionmentioning
confidence: 99%