56B3 [derivative of CRL-11117] (ATCC® CRL-2542)

Organism: Mus musculus, mouse  /  Cell Type: embryonic stem cell  / 

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Organism Mus musculus, mouse
Cell Type embryonic stem cell
Product Format frozen
Culture Properties adherent
Biosafety Level 1
Age embryo
Strain 129/Sv+c/+p
Applications
These cell lines were used to produce mutant mice with germ line transmission for lck disruption.
Comments
The replacement-type vector (PmlckBSNeo2.3) was introduced into D3 embryonic stem (ES) cells by electroporation to disrupt the lck locus.
Two cell lines, 59B5 (ATCC-CRL-11115) and 56B3 (ATCC-CRL-11117), were generated that were deficient for the lck gene.
These cell lines were used to produce mutant mice with germ line transmission for lck disruption.
Heterozygous mice were inbred to obtain mice homozygous for the disrupted lck gene.
A culture submitted to the ATCC as CRL-11117 in September of 1992 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline.
The cured cell line is available as CRL-2542. The original patent deposit is available as CRL-11117.
The line should be grown on feeder layers of mitomycin C treated (0.01 mg/ml for 90 minutes) primary mouse embryonic fibroblasts or STO cells (see ATCC CRL-1503 or ATCC 56-X.2, MITC-STO cells).
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with 0.1 mM 2-mercaptoethanol and 10mM HEPES, 85%; fetal bovine serum, 15%
Subculturing
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: 3 times per week
Remove medium and rinse the monolayer with fresh 0.25% trypsin, 0.03% EDTA solution. Remove the trypsin and incubate at 37°C until the cells detach (approximately 10 minutes). Add fresh medium, aspirate and dispense onto fresh feeder layer cultures.
Culture Conditions
Temperature: 37.0°C
Name of Depositor TW Mak
References

Mak TW. Mouse having a disrupted lck gene. US Patent 5,625,122 dated Apr 29 1997

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Mak TW. Mouse having a disrupted lck gene. US Patent 5,625,122 dated Apr 29 1997