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A9 cells are a fibroblast-like cell line derived from mouse adipose tissue. They were established as a subclone of the L929 parent strain by W. R. Earle in 1940. The parent strain was obtained from normal subcutaneous areolar and adipose tissue of a male C3H/An mouse.
A notable feature of these cells is that they express adenosine phosphoribosyl transferase (APRT) and hypoxanthine phosphoribosyl transferase (HPRT), denoted as APRT+ and HPRT+. These cells have been valuable in virus studies, particularly involving pseudorabies virus (PRV), vesicular stomatitis virus (VSV) of the Indiana strain, and herpes simplex virus (HSV).
A9 cells' sensitivity and response to these viruses have made them useful for studying viral replication, pathogenesis, and potential antiviral treatments. In immunology, A9 cells are used in various research areas. They are a valuable model for studying immune responses, antibody production, monoclonal antibody generation, and hybridoma technology.
Due to their rapid proliferation (doubling time of approximately 24 hours), A9 cells provide a sufficient cell supply for experiments and downstream applications. A9 cells have a fibroblast-like morphology and adhere to the culture substrate. Categorized as animal cells and belonging to the hybridoma cell type, A9 cells were formed by fusing B lymphocytes from Mus musculus (mouse) with myeloma cells from the same species.
This unique combination allows A9 cells to exhibit properties of both B lymphocytes and myeloma cells. Overall, A9 cells are a well-established fibroblast-like cell line utilized for studying viral infections, especially PRV, VSV, and HSV, and in immunology.
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- Antigen expression: H-2k
- Tumorigenic: Yes, in nude mice.
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- 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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