tcel183 PA317

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Product Description

Cells shipped from the ATCC have been selected in medium containing 0.03mM Hypoxanthine, 0.001mM amethopterin(Methotrexate), and 0.02mM Thymidine prior to freezing, and should be grown in HT medium for 4 days after receipt. After this, the cells are stable for at least 1 month in the absence of further selection. Introduction of retroviral vectors into these cells, by infection or by transfection, results in production of retrovirus virions with an amphotropic host range that are capable of infecting cells of many mammalian species. The percentage of PA317 cells that are capable of packaging retroviral vectors decreases slowly with continued passaging of the cell line, presumably due to the loss of the transfected DNA used to create the line. Brief selection(about 5 days) in medium containing 0.03mM Hypoxanthine, 0.001mM amethopterin(Methotrexate), and 0.02mM Thymidine will select for cells that retain the packaging function. After selection, the cells should be grown in HT medium(0.03mM Hypoxanthine, 0.02mM Thymidine) for 4 days to dilute any residual amethopterin.

Information

ApplicationVirions produced by this line have been used successfully to transfer genes into humans. Packaging line for retroviruses.
TissueMouse embryo
Morphologyfibroblast
Growth Propertiesadherent

Specifications

Complete Growth MediumDMEM (PM150210)+10% FBS (164210-500)+1% P/S (PB180120)
Subcultivation Ratio1:2-1:4
Medium Renewalevery 2 to 3 days
CryopreservationFreeze medium: 60% Basal medium+30% FBS+10% DMSO Storage temperature: Liquid nitrogen vapor phase
Culture ConditionsAtmosphere: Air, 95%; CO2, 5% Temperature: 37℃
DurationVirions produced by this line have been used successfully to transfer genes into humans. Packaging line for retroviruses.

Misc Information

SubculturingRemove and discard culture medium. Briefly rinse the cell layer with DPBS solution to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 2 to 3 minutes). Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels.
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