Instructions on Preparation and Shipping of Frozen Hybridoma Cells to GenScript for Antibody Production

Preparation and Shipping of Frozen Cells

Only cryo-preserve rapidly growing and dividing hybridomas (log phase hybridomas) that show more than 90% viability. If possible, send duplicate samples of each cell line.

• Under sterile conditions, harvest log-phase hybridoma cells in a centrifuge tube and perform a cell count using a hemacytometer and trypan blue.
• Centrifuge cells in a centrifuge tube at 200 g (or 1,000 rpm) for five minutes at room temperature.
• Carefully remove the medium and re-suspend the cell pellet in freezing medium (DMEM with 90% fetal bovine serum and 10% DMSO) to a final cell density of 5-10 x10⁶ cells per ml.
• Aliquot into cryovials at 1.0 ml/vial.
• Place the cryovials in cryo freezing containers or other suitable containers and place in freezer overnight at -80°C.
• The next day, transfer the cryo-vials to a liquid nitrogen tank. Alternatively, the frozen cells can be shipped overnight if packed in a sufficient amount of dry ice. Wear gloves and a facemask when working with liquid nitrogen tanks or frozen cryo-vials.

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