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L929-GFP
Tissue
Connective tissue
Synonyms
L929/GL50

Documents

L929-GFP

Cell line · Product number 305956

L-929-GFP cells are a fluorescently labeled derivative of the murine L-929 fibroblast cell line, which was originally established from subcutaneous connec...

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01General information

Description

L-929-GFP cells are a fluorescently labeled derivative of the murine L-929 fibroblast cell line, which was originally established from subcutaneous connective tissue of an adult mouse. The parental L-929 line is one of the most extensively used mouse fibroblast models in biomedical research and is characterized by its adherent growth, spindle-shaped morphology, and robust proliferative capacity. L-929 cells are widely utilized in studies of cytotoxicity, inflammation, extracellular matrix biology, and host–pathogen interactions, and they are also commonly employed for the production and bioassay of cytokines such as tumor necrosis factor-α (TNF-α).

The stable expression of green fluorescent protein (GFP) in L-929-GFP cells enables direct visualization and quantitative tracking of fibroblast behavior in real time. These cells are particularly useful for fluorescence-based applications including migration assays, co-culture experiments, tissue engineering studies, and live-cell imaging. L-929-GFP cells retain the core biological characteristics of the parental fibroblast line while providing enhanced utility for monitoring cell localization, proliferation, and interactions within complex cellular environments. Consequently, they serve as a versatile model for investigating stromal cell dynamics, wound healing processes, biomaterial compatibility, and immune-mediated cytotoxic responses.

Genetic modification: Stably modified by replication-incompetent lentiviral transduction to express the ZsGreen1 green fluorescent protein reporter; maintained as a polyclonal population under puromycin selection (1–5 µg/mL). S1/BSL-1 containment.

Tissue
Connective tissue
Synonyms
  • L929/GL50

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This cell line contains a stably integrated ZsGreen1 green fluorescent protein reporter introduced via replication-incompetent lentiviral transduction. The resulting polyclonal cell population was maintained under puromycin selection (1–5 µg/mL). S1 containment is required. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Cell type
Fibroblast

04Identifiers

Citation
L929-GFP (Cytion catalog number 305956)
NCBI TaxID
10090
Cellosaurus
CVCL_E2Z7

05Expression and susceptibility

Antigen expression
ZsGreen1 (green fluorescent protein)

06Handling and culture

Culture medium
DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)
Incubation atmosphere

37°C, 5% CO2, humidified atmosphere.

Supplements
Supplement the medium with 10% 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.
Seeding density
1 to 3 x 104 cells/cm2
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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

  7. Follow the procedure described under Post-Thaw Recovery

07Quality control and cryopreservation

Freeze medium
As a cryopreservation medium, we use complete growth medium + 10% DMSO for adequate post-thaw viability.
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
100 days