Lx-2

01Description

02General information

Description

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.

Tissue

Liver

Synonyms

• Lieming xu-2

03Classification

Organism

Human

Sex

Male

Growth properties

Adherent

Biosafety level

1

04Characteristics

Morphology

Epithelial

Cell type

Hepatic Stellate cells

05Identifiers

Citation

Lx-2 (Cytion catalog number 305039)

NCBI TaxID

9606

Cellosaurus

CVCL_5792

06Handling and culture

Culture medium

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)

Incubation atmosphere

37°C, 5% CO₂, humidified atmosphere.

Supplements

Supplement the medium with 2% FBS

Dissociation reagent

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.

Fluid renewal

2 to 3 times per week

Thawing and culturing

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

Freeze medium

As a cryopreservation medium, we 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.

Storage conditions

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Shipping conditions

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

08Donor

Donor age

Age unspecified

09Legal and safety

Safety precautions

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

Warranty

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.

Material Transfer Agreement required

Yes

10Product details

Composition table