| Applications |
PG13/LN c8 cells produce a retroviral vector (LN) that transduces the neo gene and can infect cells from many mammalian species (other than mice). The line was derived from PG13 (see ATCC CRL-10686). Although virus production has not been observed, there is a possibility that the cells will produce a virus similar to GaLV (a moderate risk oncogenic virus) and Biosafety Level 2 or higher precautions should be taken when using this cell line. |
| Comments |
PG13/LN c8 cells produce a retroviral vector (LN) that transduces the neo gene and can infect cells from many mammalian species (other than mice). The line was derived from PG13 (see ATCC CRL-10686). Although virus production has not been observed, there is a possibility that the cells will produce a virus similar to GaLV (a moderate risk oncogenic virus) and Biosafety Level 2 or higher precautions should be taken when using this cell line. |
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which 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: 1:4
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| References |
Miller AD, et al. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J. Virol. 65: 2220-2224, 1991. PubMed: 1850008
Miller AD, Rosman GJ. Improved retroviral vectors for gene transfer and expression. BioTechniques 7: 980-990, 1989. PubMed: 2631796
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
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