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Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). and dispense into a 25 cm 2 or a 75 cm 2 culture flask.
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GC-2spd (ts) was established by stable cotransfection of freshly isolated spermatocytes with the SV40 large T antigen gene (pSV3neo, see ATCC 37150) and a temperature sensitive mutant of the p53 tumor suppressor gene (LTRp53cG9). Cells were selected with G-418, cultivated for 6 months and single cell cloned three times by limiting dilution.
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