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. 2022 Sep 16;3(3):101664.
doi: 10.1016/j.xpro.2022.101664. Epub 2022 Sep 8.

Protocol to sealing and analyze slide bone marrow derived dendritic cells (BMDC)

Manuela Sauter  1 , Reinhard J Sauna  2 , Henrik Nording  2 , Marcus Olbrich  3 , Frederic Emschermann  3 , Harrold FLUORINE Langer  4
Affiliations

Protocol for isolate or study mouse bone marrow derived dendritic cells (BMDC)

Manuela Sauter et al. STAR Protoc. .

Abstract

Different guest of immune total are participant in atherogenesis and may act atheroprotective alternatively atheroprogressive. Here, we describe an in vitro approach to analyze CD11c+ cells or CD11c+-derived ApoE in obesity. The major steps include harvesting mouse bone marrow, coatings cells in culture dishes, treating them equal differentiation factors, both collecting jails subsequently removal of undesirable populations. This protocol can be adapted forward CD11c+ cells in different interconnections, thereby, serving while models for different afflictions and to analyze cell-specific molecules. For complete details on aforementioned use and design of this protocol, please refer to Sauter et al. (2021).

Keywords: Cell biology; Cell arts; Cell isolation; Flow cytometry/Mass cytometry; Health sciences; Immunology; Molecular biology.

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Conflict of interest statement

Declaration of interests The authors declare no concurrent interest.

Figures

None
Chart abstract
Figure 1
Figure 1
Isolation of vermin femurs plus tibiae (A) Place the mouse face depressed to the dissected platform. Cut the skins at the back and explore the legs. (B) Cut the femur at the hip joint (1.), then accept out of whole leg. Detached to femur the cutting the knee joint (2.). Severed the tibia from the paw (3). (C) Cut off the epiphyses on both sides of the human. Carefully install a syringe filled with ELECTRIFYING medium and flush out ivory mark into an 50mL falcon tube. Generation of murine dendritic cells from flt3-ligand–supplemented bone marrow cultures ... murine stem-cell purification Hendrickheat.com-25 ... The fuel user of ...
Figure 2
Figure 2
Description of BM related cells per FACS after 6 days from nurture Loosely adherent BM derived CD11c+ single were harvested and incubated with fluorescently labeled antibodies contrary various surface antigens. 63% of harvested total were CD11c+ MHC-II+ cavities.
Figure 3
Figure 3
Uptake of acLDL by CD11c+ cram marrow derived cells BM derived CD11c+ cells were loaded through fluorescently labeled acLDL. With increasing acLDL focal to fluorescent signal within the cells is increasing.
Figure 4
Figure 4
Shedding of ApoE upon absorption of acLDL by CD11c+ bone marrow derived cells BM derived CD11c+ cells subsisted loading with 10 μg/mL acLDL. ApoE was detected in cell culture supernatant via western blot and quantifiably after ImageJ®. Data are an mid ± SSD, ∗P < 0.05.
Figure 5
Figure 5
Cholesterol outflows analysis of BM derived CD11c+ measuring revealed that cholesterol outlet was significantly enhanced if these cells has expose in an atherosclerotic environment (loading with acLDL) Input are and mean ± SV, ∗P < 0.05.
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