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LX-2 is a human hepatic stellate cell line that has become a standard model for studying liver fibrosis. This cell line was immortalized from primary human hepatic stellate cells, retaining many of the in vivo characteristics necessary for the study of stellate cell activation, interaction with other liver cell types, and response to inflammatory signals. LX-2 cells are particularly noted for their utility in research focused on the pathogenesis of liver fibrosis and the evaluation of anti-fibrotic drugs. They express a variety of markers relevant to stellate cell function and fibrogenesis, including alpha-smooth muscle actin (α-SMA), glial fibrillary acidic protein (GFAP), and type I collagen.
The cell line offers an advantageous model due to its stable phenotype and responsiveness to cytokines and growth factors typically involved in liver disease processes. LX-2 cells are used to examine the cellular and molecular mechanisms underlying liver fibrosis, including the role of stellate cells in extracellular matrix deposition and the modulation of these processes by therapeutic agents. These cells provide a reproducible and controlled in vitro environment that supports high-throughput screening and mechanistic studies, making them valuable for both basic research and pharmaceutical development targeting liver diseases.
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- 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 2% 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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