36.5 (ATCC® CRL-11116)

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

Organism Mus musculus, mouse
Cell Type pluripotent embryonic stem cell
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Age embryo, blastocyst
Strain 129/Sv+c/+p
Applications
This line was derived by inserting a construct containing a disrupted murine Lyt-2 (CD8) gene.
After selection for neomycin resistance and presence of Lyt-2 sequences upstream of the insertion, the 36.5 cell line was established.
This line has been used to produce mice deficient in expression of CD8.
The cells can be maintained in the undifferentiated state by frequent subculture on feeder layers of irradiated (12000 rads) or mitomycin C treated (0.01 mg/ml for 90 minutes) primary mouse embryonic fibroblasts or STO cells.
Derivation
This line was derived by inserting a construct containing a disrupted murine Lyt-2 (CD8) gene.
Comments
This line was derived by inserting a construct containing a disrupted murine Lyt-2 (CD8) gene.
The Lyt-2 genes was disrupted by insertion of a plasmid containing a neomycin resistance gene into the first exon of the Lyt-2 gene.
After selection for neomycin resistance and presence of Lyt-2 sequences upstream of the insertion, the 36.5 cell line was established.
This line has been used to produce mice deficient in expression of CD8.
The cells can be maintained in the undifferentiated state by frequent subculture on feeder layers of mitomycin C treated STO cells (MITC-STO ATCC 56-X.2).
(see 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, 85%; fetal bovine serum, 15%
Subculturing
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium and rinse the monolayer with fresh 0.25% trypsin, 0.03% EDTA solution. Remove the trypsin and incubate at 37C until the cells detach (approximately 10 minutes). Add fresh medium, aspirate and dispense onto fresh feeder layer cultures.
Cryopreservation
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37.0°C
Name of Depositor Ontario Cancer Institute
References

Mak TW. Mutant mouse lacking CD8 surface marker. US Patent 5,530,178 dated Jun 25 1996

Basic Documentation
References

Mak TW. Mutant mouse lacking CD8 surface marker. US Patent 5,530,178 dated Jun 25 1996