LX-2 Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	LX-2 is an immortalized human hepatic stellate cell line derived from primary stellate cells isolated from normal liver tissue and selected for stable growth under reduced-serum conditions. The line retains core features of activated hepatic stellate cells, including fibrogenic marker expression, retinoid-associated functions, extracellular-matrix regulation, and cytokine-responsive signaling. These properties support reproducible in vitro studies of liver fibrosis, stellate-cell activation, matrix remodeling, metabolic stress, and antifibrotic intervention.
Cell Type	Human hepatic stellate cell
Tissue of Origin	Liver
Donor Status	Derived from primary hepatic stellate cells isolated from a normal human donor
Sex	Male
Age at Sampling	Not reported
Morphology	Adherent, spindle-shaped cells with an activated stellate-cell / myofibroblast-like phenotype
Immortalization Status	Continuous / immortalized. The line was selected under low-serum conditions; integration of SV40 large T antigen has been reported for the established cell line.
Product Format	Frozen
Growth Mode	Adherent
Applications	1. Hepatic stellate-cell activation and liver-fibrosis research 2. Extracellular-matrix deposition and remodeling studies 3. TGF-β- and PDGF-responsive signaling assays 4. Collagen-expression and fibrogenic gene-regulation studies 5. Retinoid metabolism and lipid-droplet biology 6. Oxidative, metabolic, inflammatory, and lipotoxic stress models 7. Antifibrotic compound screening and mechanism-of-action studies 8. Transfection-based functional genomics and pathway analysis
Storage Conditions	Store in the vapor phase of liquid nitrogen for long-term preservation
Characteristics
Hepatic Stellate-Cell Phenotype	Retains key functional features of activated human hepatic stellate cells, including cytokine-responsive signaling, retinoid-associated biology, fibrogenic activity, and neuronal-lineage gene expression described for this cell type.
Marker Expression	Expresses α-smooth muscle actin (α-SMA), vimentin, and glial fibrillary acidic protein (GFAP)
Fibrosis-Related Receptors	Expresses platelet-derived growth factor receptor-β (PDGFR-β), the long-form leptin receptor (Ob-RL), and discoidin domain receptor 2 (DDR2)
Matrix-Remodeling Proteins	Expresses matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinases-2 (TIMP-2), and membrane-type 1 matrix metalloproteinase (MT1-MMP). TIMP-1 expression is lower than in primary activated stellate cells in the original characterization.
Collagen-Related Expression	Expresses α1(I) procollagen and heat-shock protein 47 (HSP47) transcripts
Growth-Factor Response	PDGF stimulates proliferation, while TGF-β1 increases α1(I) procollagen transcript expression under the reported assay conditions
Retinoid Phenotype	Displays retinoid-associated features characteristic of hepatic stellate cells and retains reported capacity for retinol uptake and esterification
Transfection Suitability	Reported to be highly transfectable, supporting transient gene-expression and pathway-perturbation studies
Mycoplasma Test	Negative
Culture Conditions and Handling
Culture Medium	Routine expansion: high-glucose DMEM supplemented with 2% fetal bovine serum, 1× L-glutamine, and 1× penicillin–streptomycin. For initial post-thaw recovery, use the same basal medium with 10% fetal bovine serum before changing to the reduced-serum expansion medium.
Culture Conditions	37°C in a humidified atmosphere containing 5% CO2
Culture Vessel	Standard tissue-culture-treated vessels; no matrix coating is required in the referenced protocol
Medium Renewal	Replace the medium every 2-3 days
Passage Endpoint	Subculture at approximately 80% confluence; avoid prolonged over-confluence when comparing fibrogenic responses across experiments
Subculturing	1. Remove the spent medium and rinse the monolayer with calcium- and magnesium-free balanced salt solution. 2. Add 3–5 mL of a gentle cell-dissociation reagent or trypsin to a T-75 flask and incubate at 37°C for approximately 3-5 minutes. 3. Confirm detachment microscopically and avoid unnecessary exposure to the dissociation reagent. 4. Add approximately 8 mL of complete 2% serum medium, collect the cells, and centrifuge at 300 × g for 3–5 minutes. 5. Resuspend gently without vortexing and transfer the required aliquot to fresh culture vessels. 6. Return the cultures to 37°C and 5% CO2.
Subcultivation Ratio	A split ratio of 1:3 to 1:6 is reported for routine expansion
Thawing and Recovery	Thaw the vial rapidly in a 37°C water bath and disinfect the exterior before opening. Transfer the suspension without vortexing and slowly add approximately 9 mL of pre-warmed recovery medium containing 10% fetal bovine serum. Centrifuge at 300 × g for 2-3 minutes, resuspend gently, seed into a T-75 flask, and incubate overnight. Replace with the 2% serum expansion medium on the following day.
Cryopreservation	A reported freezing formulation consists of complete growth medium adjusted to 20% fetal bovine serum and 10% DMSO. Freeze at a controlled rate before transfer to liquid nitrogen.
Handling Notes	Use aseptic technique and gentle pipetting; do not vortex the cells. Because proliferation and fibrogenic readouts vary with serum concentration, passage history, and confluence, standardize these variables within each experiment. Confirm identity and contamination status periodically, and document the medium formulation and passage number used for each study.

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