Cell Line: L-929

OrganismMouse (Mus musculus), 100-day-old male, C3H/An
Source/Tissue of origin
Subcutaneous connective tissue, Adipose, Areolar
Cell typeConnective tissue fibroblasts
Growth modeAdherent, monolayer
Doubling time
20-24 hours
Life spanInfinite, continuous culture
Karyotype Modal chromosome number = 66; range = 65 to 68
Tumorigenic Tumorigenic in immunosuppressed mice
Drug resistance 
Biosafety Level1 (dsmz/atcc) (May differ in your country, follow the guidelines of your institute/country)
Gene Expression 
Applications 3D cell culture, toxicity testing
Related cell lines 

Cell line isolation information

The cell line L-929 was established from the normal subcutaneous areolar and adipose tissue of a male C3H/An mouse.

Recommended propagation method

Culture medium:
1. 90% RPMI 1640 + 10% h.i. FBS (dsmz)
2. 90% Eagle’s Minimum Essential Medium + 10% horse serum (atcc)

Culture conditions: Humidified incubator, 95% air + 5% CO2, 37°C.
Split ratio: 1:5 to 1:10
Split frequency: 2-3 times a week
Harvesting time: Confluent culture
Split Method: Trypsin (do not use trypsin/EDTA) (dsmz)
Seeding density: 2.0-3.0 x 106 cells/80 cm2

Recommended Cryopreservation Method

Freeze medium:
1. 70% complete medium + 20% FBS  + 10% DMSO (dsmz)
2. Complete growth medium supplemented with 5% (v/v) DMSO (atcc)

Storage temperature: Liquid nitrogen vapor phase

Cell density:



  • Sanford et al., 1948. The growth in vitro of single isolated tissue cells. J Natl Cancer Inst. 9(3), 229-246.  PMID-18105872

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