bmdm cell line tlr2  (BEI Resources)

Journal: Regenerative Therapy

Article Title: In vitro and in vivo study of concentrated growth factor (CGF) mediating macrophage polarization in bone defect repair

doi: 10.1016/j.reth.2025.04.013

Figure Lengend Snippet: Surface characteristics of C GF and its effect on macrophages. A: CGF and its three-dimensional structure under electron microscopy; The proliferation of BMDM treated with different concentrations of CGF; B: ELISA method is used to detect the secretion of IL-1 β 、TNF- α 、VEGF-A、IL-10 after CGF acts on M0 and M1 macrophages; C: Western blot detection of protein expression of Arg-1 and iNOS after CGF acts on M0 and M1 macrophages; D: Western blot detection of JAK/STAT pathway related protein expression after CGF acts on M0 and M1 macrophages, respectively (∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001. #, P < 0.05; ##, P < 0.01; ###, P < 0.001; ####, P < 0.0001. # compared with the control group, ns, P > 0.05,one-way ANOVA).

Article Snippet: BMDM macrophage cell lines (Procell Life Science & Technology Co., Ltd.) were used for in vitro studies.

Techniques: Electron Microscopy, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: A Novel Mechanism of Macrophage Activation by the Natural Yolkin Polypeptide Complex from Egg Yolk

doi: 10.3390/ijms23063125

Figure Lengend Snippet: Effect of yolkin on viability and proliferation of bone marrow-derived macrophages (BMDM). The cells (1 × 10 5 /mL) were seeded in 96-well plates in DMEM + 10% FBS and incubated overnight at 37 °C. Next, the cells were exposed to yolkin (10, 100 and 150 µg/mL) for 24 and 48 h. The cell proliferation activity of yolkin was assessed with an MTT assay. Non-stimulated cells were used as a negative control. The results represent three to five independent experiments and data are presented as median ± min–max. A one-sample t -test was used to examine the mean differences between samples and control (100). * p ≤ 0.05, *** p ≤ 0.0001.

Article Snippet: The murine bone marrow-derived macrophages of the BMDM cell line (Bei Resources) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS, antibiotics (penicillin, streptomycin and gentamycin) and 3% L-glutamine.

Techniques: Derivative Assay, Incubation, Activity Assay, MTT Assay, Negative Control, Control

Journal: Journal of Innate Immunity

Article Title: HSV-1/TLR9-Mediated IFNβ and TNFα Induction Is Mal-Dependent in Macrophages

doi: 10.1159/000504542

Figure Lengend Snippet: a, b Knockout of Mal abrogates the production of IFNβ and TNFα in murine macrophages. WT and Mal-deficient (MalKO) BMDM cells were stimulated with the TLR9 ligand CpG-C (5 µM) for 4 h. Thereafter, total RNA was isolated and reverse transcribed. The relative expression of Ifnb1 and Tnf was assayed with the use of qPCR (2−(ΔΔCT)). Relative expression values were normalized to the Hprt1 reference gene. c, d Level of biologically active type I IFNs and TNFα in BMDM cells ­stimulated with the TLR9 ligand. WT and MalKO BMDM cells were stimulated with CpG-C (5 µM) for 24 h. Subsequently, the level of IFNα/β or TNFα in culture medium was measured with the use of bioassay and ELISA, respectively. The results are representative of at least three independent experiments. *p < 0.001 (unpaired Student t test).

Article Snippet: Cell Culture and Reagents Immortalized BMDM cell lines from wild-type (WT) and Mal–/– mice were obtained from Bei Resources.

Techniques: Knock-Out, Isolation, Reverse Transcription, Expressing, Bioassay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Innate Immunity

Article Title: HSV-1/TLR9-Mediated IFNβ and TNFα Induction Is Mal-Dependent in Macrophages

doi: 10.1159/000504542

Figure Lengend Snippet: TLR9-induced activation of NF-κB in macrophages is Mal-dependent. a Translocation of the NF-κB transcription factor in BMDM cells treated with TLR9 ligand. WT and MalKO BMDM cells were stimulated with CpG-C (5 µM) for 2 h. The nuclear fractions were then isolated and incubated with specific, fluorescently labeled oligonucleotide probes to bind the NF-κB factor. The samples were electrophoresed on a polyacrylamide gel. Visualization was perform-ed using the Odyssey CLx Imaging System (LI-COR). The results presented are representative of at least three independent experiments. b–d Effect of inhibition of NF-κB translocation on Ifnb1, Tnf expression, and biologically active type I IFN release in BMDM cells. Cells were stimulated with CpG-C (5 μM) for 4 h, in the presence or absence of JSH-23 (10 μM), the NF-κB nuclear translocation inhibitor, added 1 h prior to stimulation. Next, total RNA was isolated and relative expression of Ifnb1 (b) or Tnf (c) was assayed with qPCR (2−(ΔΔCT)) using Hprt1 for normalization. Cells were stimulated with CpG-C at 5 μM for 16 h in the presence or absence of JSH-23 (10 μM) added 1 h prior to stimulation. Subsequently, the level of type I interferons in supernatants was determined by a biological method (d). Data are compiled from at least three independent experiments, each experiment being performed in duplicate (mean ± SD). *p < 0.001.

Article Snippet: Cell Culture and Reagents Immortalized BMDM cell lines from wild-type (WT) and Mal–/– mice were obtained from Bei Resources.

Techniques: Activation Assay, Translocation Assay, Isolation, Incubation, Labeling, Imaging, Inhibition, Expressing