NCTC clone 929 [L cell, L-929, derivative of Strain L]
Depositors:
WR Earle
Biosafety Level:
1
Shipped:
frozen
Medium & Serum:
See Propagation
Growth Properties:
adherent
Organism:
Mus musculus
Morphology:
fibroblast
Source:
Tissue: subcutaneous connective tissue; areolar and adipose Strain: C3H/An
Permits/Forms:
In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimay responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
modal chromosome number = 66; range = 65 to 68. There were approximay 20 to 30 marker chromosomes present in each metaphase spread. A high percentage of those markers were common to most analyzed cells. A long metacentric chromosome with secondary constriction was noted in 77/100 cells.
Age:
100 days
Gender:
male
Comments:
NCTC clone 929 （Connective tissue, mouse） Clone of strain L was derived in March, 1948. Strain L was one of the first cell strains to be established in continuous culture, and clone 929 was the first cloned strain developed. The parent L strain was derived from normal subcutaneous areolar and adipose tissue of a 100-day-old male C3H/An mouse. Clone 929 was established （by the capillary technique for single cell isolation） from the 95th subculture generation of the parent strain. Tested and found negative for ectromelia virus （mousepox）.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle’s Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: horse serum to a final concentration of 10%. Atmosphere: air, 95%; carbon dioxide （CO2）, 5% Temperature: 37.0°C
Subculturing:
Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% （w/v） Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed （usually within 5 to 15 minutes）. Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
Preservation:
Freeze medium: Complete growth medium supplemented with 5% （v/v） DMSO Storage temperature: liquid nitrogen vapor phase
Related Products:
Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2003 derivative:ATCC CCL-1.1 derivative:ATCC CCL-1.2 derivative:ATCC CCL-1.3 derivative:ATCC CCL-1.4
References:
3: Kazazian HH Jr., et al. Restriction site polymorphism in the phosphoglycerate kinase gene on the X chromosome. Hum. Genet. 66: 217-219, 1984. PubMed: 6325324 21373: Fisher EM, et al. Homologous ribosomal protein genes on the human X and Y chromosomes: escape from X inactivation and possible implications for Turner syndrome. Cell 63: 1205-1218, 1990. PubMed: 2124517 21404: Sanford KK, et al. The growth in vitro of single isolated tissue cells. J. Natl. Cancer Inst. 9: 229-246, 1948. 21405: Sugarman BJ, et al. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science 230: 943-945, 1985. PubMed: 3933111 21469: ASTM International Standard Practice for Direct Contact Cell Culture Evaluation of Materials for Medical Devices. West Conshohocken, PA:ASTM International;ASTM Standard Test Method F 0813-07. 21470: ASTM International Standard Test Method for Agar Diffusion Cell Culture Screening for Cytotoxicity. West Conshohocken, PA:ASTM International;ASTM Standard Test Method F 0895-84 （Reapproved 2006）. 21606: U.S. Pharmacopeia General Chapters: <87> Biological Reactivity Tests, in vitro. Rockville, MD: U.S. Pharmacopeia; USP USP34-NF29, 2011 23579: Westfall BB, et al. The glycogen content of cell suspensions prepared from massive tissue culture: comparison of cells derived from mouse connective tissue and mouse liver. J. Natl. Cancer Inst. 14: 655-664, 1953. PubMed: 13233820 25770: Earle WR, et al. Production of malignancy in vitro. IV. The mouse fibroblast cultures and changes seen in the living cells. J. Natl. Cancer Inst. 4: 165-212, 1943. 25879: Earle WR, et al. The influence of inoculum size on proliferation in tissue cultures. J. Natl. Cancer Inst. 12: 133-153, 1951. PubMed: 14874126 25880: Sanford KK, et al. The tumor-producing capacity of strain L mouse cells after 10 years in vitro. Cancer Res. 16: 162-166, 1956. PubMed: 13293658 25882: Westfall BB, et al. The arginase and rhodanese activities of certain cell strains after long cultivation in vitro. J. Biophys. Biochem. Cytol. 4: 567-570, 1958. PubMed: 13587550 29223: Papkoff J. Regulation of complexed and free catenin pools by distinct mechanisms. J. Biol. Chem. 272: 4536-4543, 1997. PubMed: 9020180 32283: Hu SX, et al. Development of an adenovirus vector with tetracycline-regulatable human tumor necrosis factor alpha gene expression. Cancer Res. 57: 3339-3343, 1997. PubMed: 9269991 33114: Yasin B, et al. Susceptibility of Chlamydia trachomatis to protegrins and defensins. Infect. Immun. 64: 709-713, 1996. PubMed: 8641770 92346: Biological evaluation of medical devices. Part 5: Tests for in vitro cytotoxicity. Sydney, NSW, Australia:Standards Australia;Standards Australia AS ISO 10993.5-2002. 92380: Plastics collapsible containers for human blood and blood components–Part 1: Conventional containers. Annex C. Biological tests.. Geneva （Switzerland）:International Organization for Standardization/ANSI;ISO ISO 3826-1:2003. 92382: Dentistry–Preclinical evaluation of biocompatibility of medical devices used in dentistry–Test methods for dental materials. Geneva （Switzerland）:International Organization for Standardization/ANSI;ISO ISO 7405:1997. 92389: Biological evaluation of medical devices–Part 5: Tests for in vitro cytotoxicity. Geneva （Switzerland）:International Organization for Standardization/ANSI;ISO ISO 10993-5:1999. 92404: Plastic containers for intravenous injection. Geneva （Switzerland）:International Organization for Standardization/ANSI;ISO ISO 15747:2003.
