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DH-82 cells, derived from the malignant histiocytosis of a ten-year-old male Golden Retriever, are a cornerstone in the study of canine immunology and related diseases.
These cells exhibit a macrophage-like morphology, mirroring the key functions of human macrophages, thereby providing a relevant model for investigating various aspects of canine health, particularly immune system-related conditions.
A defining characteristic of DH-82 cells is their capability to phagocytize latex particles, an essential function of macrophages responsible for the elimination of foreign substances in the body. This property positions DH-82 cells as a robust tool for delving into the immune responses of dogs, especially in the face of infections and inflammatory diseases. The expression of Fc gamma receptors in DH-82 cells is a notable trait.
These receptors are integral to immune responses, as they bind to antibodies and facilitate the phagocytosis of antibody-coated pathogens or particles. This makes DH-82 cells particularly valuable in studies focusing on immune responses and antibody-dependent cellular cytotoxicity (ADCC). In contrast, DH-82 cells do not express Fc mu and C3b receptors.
The absence of Fc mu receptors, typically found on B cells and involved in antigen presentation, and C3b receptors, which bind to complement proteins in immune responses, provides a controlled setting for examining specific immune mechanisms that might be influenced by these receptors.
Additionally, DH-82 cells are non-producers of IL-1, a pivotal cytokine in inflammatory responses. This feature offers a unique perspective for investigating the role of IL-1 in various biological processes and understanding IL-1-mediated diseases.
In the realm of infectious diseases, DH-82 cells have proven particularly useful in studying canine monocytic ehrlichiosis (CME), a tick-borne illness caused by Ehrlichia canis.
The cells provide a conducive environment for the bacterium's growth, aiding in the exploration of the disease's development and potential treatments. The doubling time of DH-82 cells, approximately 26 hours, is also a critical aspect in their use, influencing experimental design and the interpretation of results.
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- cultureMedium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
- supplements: Supplement the medium with 10% FBS and 1% NEAA
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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