GP+envAM-12 (ATCC® CRL-9641)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast  /  Tissue: embryo  / 

Organism Mus musculus, mouse
Tissue embryo
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Age embryo
Strain NIH/Swiss
Applications
It can be used to produce retroviral vectors for delivery of foreign genes into susceptible eukaryotic cells.

Derivation
The line was established by electroporation into NIH 3T3 cells of two plasmids that separately encode the the env region of a murine amphotropic MuLV and the gag, pol and other sequences needed for viral packaging.
Comments
This line is capable of packaging nucleic acids containing a psi packaging sequence into recombinant amphotropic retrovirus genomes.


Complete Growth Medium

The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002.

To make the complete growth medium, add the following components to the base medium:
• bovine calf serum to a final concentration of 10%

After 1 to 3 days, add
• 0.015 mg/ml hypoxanthine
• 0.25 mg/ml xanthine
• 0.025 mg/ml mycophenolic acid
• 0.2 mg/ml hygromycin B

Subculturing
Remove medium, add fresh 0.10% trypsin in phosphate buffered saline for 3 to 5 minutes, remove trypsin and let the culture sit at 37°C for 10 to 15 minutes. Add fresh medium, aspirate and dispense into new flasks.
Medium Renewal: Twice per week
Culture Conditions
Temperature: 37°C
Name of Depositor Trustees of Columbia University
References

Bank A, et al. Retroviral packaging cell lines and process of using same. US Patent 5,278,056 dated Jan 11 1994

Medin JA, et al. Correction in trans for Fabry disease: expression, secretion, and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector. Proc. Natl. Acad. Sci. USA 93: 7917-7922, 1996. PubMed: 8755577

Basic Documentation
References

Bank A, et al. Retroviral packaging cell lines and process of using same. US Patent 5,278,056 dated Jan 11 1994

Medin JA, et al. Correction in trans for Fabry disease: expression, secretion, and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector. Proc. Natl. Acad. Sci. USA 93: 7917-7922, 1996. PubMed: 8755577