Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes

Wagner, Martin; Koester, Helen; Deffge, Christian; Weinert, Soenke; Lauf, Johannes; Francke, Alexander; Lee, Jerry; Dullaeus, R. C. Braun-; Herold, Joerg

doi:10.3791/52347-v

Cited by 8 publications

(8 citation statements)

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“…We first tested our expectation that PODS crystals can be phagocytosed by primary murine monocytes and macrophages. Murine bone marrow-derived monocytes (BMDM) were isolated according to Wagner et al [ 16 ] from the tibias of C57BL/6 mice. Cells were cultured for 5 days before they were incubated with fresh complete medium containing PODS M-CSF or PODS GM-CSF at a ratio of 5 crystals per cell.…”

Section: Resultsmentioning

confidence: 99%

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Phagocytosed Polyhedrin-Cytokine Cocrystal Nanoparticles Provide Sustained Secretion of Bioactive Cytokines from Macrophages

et al. 2021

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Many cells possess the ability to engulf and incorporate particles by phagocytosis. This active process is characteristic of microorganisms as well as higher order species. In mammals, monocytes, macrophages, and microglia are among the so-called professional phagocytes. In addition, cells such as fibroblast and chondrocytes are classified as nonprofessional phagocytes. Professional phagocytes play important roles in both the innate and adaptive immune responses, wound healing, and tissue homeostasis. Consequently, these cells are increasingly studied as targets and vectors of therapeutic intervention to treat a range of diseases. Professional phagocytes are notoriously difficult to transfect limiting their study and manipulation. Consequently, efforts have shifted towards the development of nanoparticles to deliver a cargo to phagocytic cells via phagocytosis. However, this approach carries significant technical challenges, particularly for protein cargos. We have focused on the development of nanoscale cocrystalline protein depots, known as PODS®, that contain protein cargos, including cytokines. Here, we show that PODS are readily phagocytosed by nonprofessional as well as professional phagocytic cells and have attributes, such as highly sustained release of cargo, that suggest potential utility for the study and exploitation of phagocytic cells for drug delivery. Monocytes and macrophages that ingest PODS retain normal characteristics including a robust chemotactic response. Moreover, the PODS-cytokine cargo is secreted by the loaded cell at a level sufficient to modulate the behavior of surrounding nonphagocytic cells. The results presented here demonstrate the potential of PODS nanoparticles as a novel molecular tool for the study and manipulation of phagocytic cells and for the development of Trojan horse immunotherapy strategies to treat cancer and other diseases.

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Section: Resultsmentioning

confidence: 99%

“…Murine bone marrow-derived monocytes were isolated according to Wagner et al [ 16 ]. Briefly, the tibias of 3 C57BL/6 mice were prepared for harvest.…”

Section: Methodsmentioning

confidence: 99%

Phagocytosed Polyhedrin-Cytokine Cocrystal Nanoparticles Provide Sustained Secretion of Bioactive Cytokines from Macrophages

et al. 2021

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“…The collected cells were filtered through 40 µm cell strainers and centrifuged at 350 g for 7.5 min. Then, the bone marrow cells were resuspended in RPMI supplemented with 20 ng/mL M-CSF and plated at a density of 1 × 10 6 cells/mL in 6-well ultra-low attachment plates for differentiation into monocytes ( 49 , 50 ). For studies with human monocytes, primary blood-derived monocytes were purchased from STEMCELL.…”

Section: Methodsmentioning

confidence: 99%

A backpack-based myeloid cell therapy for multiple sclerosis

Kapate

Dunne

Kumbhojkar

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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Multiple sclerosis (MS) is an incurable autoimmune disease and is currently treated by systemic immunosuppressants with off-target side effects. Although aberrant myeloid function is often observed in MS plaques in the central nervous system (CNS), the role of myeloid cells in therapeutic intervention is currently overlooked. Here, we developed a myeloid cell-based strategy to reduce the disease burden in experimental autoimmune encephalomyelitis (EAE), a mouse model of progressive MS. We developed monocyte-adhered microparticles (“backpacks”) for activating myeloid cell phenotype to an anti-inflammatory state through localized interleukin-4 and dexamethasone signals. We demonstrate that backpack-laden monocytes infiltrated into the inflamed CNS and modulated both the local and systemic immune responses. Within the CNS, backpack-carrying monocytes regulated both the infiltrating and tissue-resident myeloid cell compartments in the spinal cord for functions related to antigen presentation and reactive species production. Treatment with backpack-monocytes also decreased the level of systemic pro-inflammatory cytokines. Additionally, backpack-laden monocytes induced modulatory effects on T H 1 and T H 17 populations in the spinal cord and blood, demonstrating cross talk between the myeloid and lymphoid arms of disease. Backpack-carrying monocytes conferred therapeutic benefit in EAE mice, as quantified by improved motor function. The use of backpack-laden monocytes offers an antigen-free, biomaterial-based approach to precisely tune cell phenotype in vivo, demonstrating the utility of myeloid cells as a therapeutic modality and target.

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“…BMMSCs and osteoclast precursors were harvested and cultured from 4–8 weeks old C57BL/6 mice BM as described. [ 40,41 ] Briefly, tibias and femurs were dissociated, scrubbed to remove any soft tissue, and transferred into ice‐cold complete media (CM) composed of minimal essential medium (MEM) α (α‐MEM) (Gibco, Thermo‐Fisher, USA) supplemented with 15% FBS (Gibco, Thermo‐Fisher, USA) and 1% penicillin‐streptomycin (Gibco, Thermo‐Fisher, USA).…”

Section: Methodsmentioning

confidence: 99%

Bioengineering Bone‐on‐a‐Chip Model Harnessing Osteoblastic and Osteoclastic Resolution

et al. 2023

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Organ-on-a-chip (OoC) systems allow the generation of microphysiological tissue models that can recapitulate key biological processes in healthy and diseased states. OoC bone models provide valuable tools to study crosscellular interactions that take place in bone-related processes. Although few bone-on-a-chip models have been proposed, structural and biological hierarchy to establish a functional unit is often lacking. Herein, a functional OoC-based 3D bone co-culture model is reported. This model comprises a highly porous β-tricalcium phosphate (TCP) based scaffold that is seeded with primary osteoblast and osteoclast precursors. This engineered construct is formed and cultured dynamically inside an OoC platform for up to 21 days and exhibits a dense extracellular matrix (ECM). Further, cultured constructs are ectopically implanted in C57BL/6 mice for 8 weeks, then histological and tartrate-resistant acid phosphatase (TRAP) analyses are carried out. These results demonstrate that both bone deposition and resorption processes are present in the bioengineered model. This study also has implications for understanding complex cellular cross-talks occurring in the bone remodeling process.

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Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes

Cited by 8 publications

References 0 publications

Phagocytosed Polyhedrin-Cytokine Cocrystal Nanoparticles Provide Sustained Secretion of Bioactive Cytokines from Macrophages

Phagocytosed Polyhedrin-Cytokine Cocrystal Nanoparticles Provide Sustained Secretion of Bioactive Cytokines from Macrophages

A backpack-based myeloid cell therapy for multiple sclerosis

Bioengineering Bone‐on‐a‐Chip Model Harnessing Osteoblastic and Osteoclastic Resolution

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