Dc2.4 cells

Manufactured by American Type Culture Collection

Sourced in United States

The DC2.4 cells are a murine (mouse) dendritic cell line derived from bone marrow. They are commonly used as an in vitro model for studying dendritic cell biology and function.

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DC2.4 dendritic cells (3 citations)

8 protocols using dc2.4 cells

Culturing Diverse Cancer Cell Lines

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CT26 cells (murine CRC cells), HCT116 cells (human colorectal cancer cells), DC2.4 cells (mouse dendritic cell line ), and HUVEC cells (human umbilical vein endothelial cells) were acquired from ATCC. These cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium at 37 °C in a humidified atmosphere with 5% CO₂. The RPMI‐1640 medium was supplemented with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin.
For the purpose of in vivo studies, BALB/C mice (female, 6–8 weeks old) were purchased from the Shanghai Wushi Experimental Animal Center (Shanghai, China). All animal studies were approved by the Institutional Animal Care and Use Committee at Fujian Medical University.

Wang L., Zhou H., Chen Q., Lin Z., Jiang C., Chen X., Chen M., Liu L., Shao L., Liu X., Pan J., Wu J., Song J., Wu J, & Zhang D. (2023). STING Agonist‐Loaded Nanoparticles Promotes Positive Regulation of Type I Interferon‐Dependent Radioimmunotherapy in Rectal Cancer. Advanced Science, 11(7), 2307858.

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Cellular Uptake of FITC-IK Nanoparticles

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DC2.4 cells (ATCC, Manassas, VA. 5×10⁵ cells/well) were seeded into 12-well plates and maintained at 37 °C in 5% CO₂ in RPMI 1640 medium supplemented with 10% FBS and 100 U/mL penicillin and 0.1 mg/mL streptomycin overnight. The medium was replaced by the same amount of FITC-IK NPs and free FITC-IK DMSO solution (40 μg/mL or 80 μg/mL, 1 mL). After 12 hr of incubation, the supernatant in each well was removed. The cells were washed 3 times with cold PBS and fixed with 4% paraformaldehyde for 20 min. Then the cells were washed 3 times again with PBS and stained with DAPI for 5min. The cellular uptake of FITC-IK NPs and free FITC-IK solution were monitored by fluorescence microscope (Olympus, Tokyo, Japan).

Hong J., Xiao X., Gao Q., Li S., Jiang B., Sun X., Ran P, & Yang P. (2019). Co-delivery of allergen epitope fragments and R848 inhibits food allergy by inducing tolerogenic dendritic cells and regulatory T cells. International Journal of Nanomedicine, 14, 7053-7064.

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Cell Line Culture Conditions

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Fl free Ce6 À Fl Ce6 in NR-MS-Ce6 Fl free Ce6 Â 100%
Cell culture B16-OVA, EMT6 and DC2.4 cells (ATCC, USA) were cultured in RPMI-1640 medium (Gibco, U.S.) supplemented with 10% fetal bovine serum (FBS) (GE Healthcare, UK), 1% sodium pyruvate (Gibco, U.S.), 1% 1 M HEPES buffer (Gibco, USA), 1% MEM non-essential amino acids (Gibco, USA) and 100 U mL À1 penicillin-streptomycin (Gibco, USA) at 37 1C, 5% CO 2 and 95% humidity. J774A.1 cells were cultured in 1Â DMEM medium (Gibco, U.S.) with the same constituents as RPMI-1640 at 37 1C, 5% CO 2 and 95% humidity.

Kang M.W.C., Liu H, & Kah J.C.Y. (2020). Innate immune activation by conditioned medium of cancer cells following combined phototherapy with photosensitizer-loaded gold nanorods. Journal of materials chemistry. B, 8(47).

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Cell Culture Protocol for Immune Cell Lines

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HEK-Blue™ mTLR2 and HEK-Blue™ Null2 cells were purchased from Invivogen and cultured in DMEM medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. DC2.4 cells were purchased from ATCC and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. Bone marrow-derived dendritic cells (BMDCs) were isolated from 6 to 8 weeks C57BL/6J mice and cultured in RPMI 1640 medium supplemented with 10% inactivated fetal bovine serum and 1% penicillin-streptomycin. 20 ng/mL GM-CSF and 10 ng/ml IL4 was used for BMDC differentiation.

Xie D., Niu Y., Mu R., Campos de Souza S., Yin X., Dong L, & Wang C. (2022). A Toll-like Receptor-Activating, Self-Adjuvant Glycan Nanocarrier. Frontiers in Chemistry, 10, 864206.

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Microparticle-Based Vaccine Formulations

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C8-HSL, N-octanoyl-DL-homoserine lactone, was purchased from Sigma-Aldrich. Polymer poly (lactic-co-glycolic) acid (PLGA) 75:25 was purchased from Evonik Industries, Alabama, and the bovine serum albumin (BSA, Cat. A3294-50G) used to formulate microparticles was purchased from Sigma-Aldrich. Dendritic cells (DC 2.4) were purchased from ATCC. DC 2.4 cells were cultivated in Dulbecco’s Modified Eagle Medium (DMEM), purchased from ATCC. Alum and MF59^® were purchased from InvivoGen, San Diego, CA. CpG adjuvant was obtained through BEI Resources, NIAID, NIH: CpG 7909 Adjuvant, NR-52393. The measles vaccine was purchased from the Serum Institute of India (SII), Pune, India. Characterization of microparticles for size, charge, and polydispersity index (PDI) were measured via Zetasizer Nano ZS (Malvern Pananalytical, MA, USA). Zika virus was received by the Centers for Disease Control and Prevention (CDC), Colorado. Measles vaccines were received from the serum institute of India. Influenza vaccine was received from BEI resources. Colonization factor antigen I (CFA/I) from E. coli. and inactive protective antigen (PA) were received from BEI resources.

Shah S.M., Joshi D., Chbib C., Roni M.A, & Uddin M.N. (2023). The Autoinducer N-Octanoyl-L-Homoserine Lactone (C8-HSL) as a Potential Adjuvant in Vaccine Formulations. Pharmaceuticals, 16(5), 713.

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Immune Cell Culture and Mouse Experiments

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Both types of mouse lymphocytes used in the experiment, namely DC2.4 cells and E.G7-OVA cells, were purchased from the American Type Culture Collection (ATCC). BALB/c mice and C57BL/6 mice (SPF, female, 6–8 weeks of age) were purchased from Beijing HFK Bioscience and Hunan SJA Laboratory Animal Co. (China), respectively. The animal experimental protocols were approved by the Laboratory Animal Welfare and Ethics Committee of the Third Military Medical University and performed according to the Guide for the Care and Use of Laboratory Animals (27 (link)); professional technicians, skilled operation of equipment, and suitable experimental conditions for the animals were ensured in order to minimize pain experienced by the mice.

Luo X., Song Z., Zeng X., Ye Y., Zheng H., Cai D., Yuan Q., Li H., Tong Y., Lu D., Liu Y., Zeng H., Yang Y., Sun H, & Zou Q. (2023). A promising self-nanoemulsifying adjuvant with plant-derived saponin D boosts immune response and exerts an anti-tumor effect. Frontiers in Immunology, 14, 1154836.

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In vitro cytotoxicity of Dex-SS-DCA nanoprobe

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MCF-7 and DC 2.4 cells (American Type Culture Collection [ATCC], Manassas, VA, USA) were incubated in full cell culture medium (ie, DMEM supplemented with 10% FBS, 100 U/mL penicillin and streptomycin [Thermo Fisher Scientific, Waltham, MA, USA]) at 37°C and in 5% CO₂. In vitro cytotoxicity of the Dex–SS–DCA nanoprobe was evaluated by assay of the cell viability. In brief, MCF-7 or DC 2.4 cells (1×10⁴ cells/well) were seeded in a 96-well plate and cocultured with the nanoprobe at polymer concentrations from 50 μg/mL to 1,000 μg/mL in full DMEM culture medium containing 10% FBS for 24 hours or 48 hours. Afterward, the cells were washed twice with fresh 1× PBS and cell viability was detected by AlamarBlue assay (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s protocol. The value for untreated cells (as blank) was taken as 100% cell viability.

Li J., Jiang B., Lin C, & Zhuang Z. (2014). Fluorescence tomographic imaging of sentinel lymph node using near-infrared emitting bioreducible dextran nanogels. International Journal of Nanomedicine, 9, 5667-5682.

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Dex-loaded PLGA Nanoparticle Transfection

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NIH/3T3 cells and DC2.4 cells were purchased from the American Type Culture Collection. The 3T3 cells and DC2.4 cells were cultured in Dulbecco’s modified Eagle’s medium containing 10% FBS at 37°C under 5% CO₂. After different concentrations of Dex-loaded PLGA nanoparticle treatment, the cell was transfected with polymer/RNP nanocomplexes. The doses of protein, sgRNA, and polymer in each well were 1, 0.5, and 8 μg, respectively. After incubation for 6 hours, the culture media containing transfection mixtures were removed and further incubated for 42 hours.

Wan T., Pan Q, & Ping Y. (2021). Microneedle-assisted genome editing: A transdermal strategy of targeting NLRP3 by CRISPR-Cas9 for synergistic therapy of inflammatory skin disorders. Science Advances, 7(11), eabe2888.

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