FRZm3 | Animal-free freezing medium

FRZm3 is a chemically defined, animal component- as well as protein-free media formulation to efficiently freeze cell lines, stem cells and spheroidal cultures.
The medium is available in two variations:

• A standard freezing medium (100303a-100ml) providing viable cell storage up to 10e7 cells/ml for daily using for cryo preservations between 0.1-2.0 ml;
• A high-performance freezing medium (100303b-10ml) providing viable cell storage up to 10e9 cells/ml. Specifically designed for immediatelly-to-use cryopresevations for assay-ready or transfection-ready approaches. Hence, it can be used as electroporation medium for mammalian cells as well.

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Application note | Characteristics

FRZm3is available in different formates:

• Standard freezing medium (100303a-100ml)
• High-performance freezing and electroporation medium (100303b-10ml)

The medium is provided as a 0.22 μm sterile filtered as well as sterility tested liquid and is free of mycoplasma.
FRZm3 contains DMSO.

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Protocol

Freezing

• Dissociate and centrifuge the cells at your standard settings;
• Optionally for aliquots for electroporation: Wash with PBS 1x once and collect the cells by centrifugation;
• Add the appropriate amount of FRZm3;
• Place the cells in a freezing container and allow gentle freezing at -80 °C for at least 10 hours.

Thawing/Electroporation

• For cryopreservation volumes >0.25 ml place the tube in a water bath at 37 °C and observe the thawing process carefully for 1-3 minutes until a small frozen piece is still visible;
• For cryopreservation volumes <0.25 ml place the tube at room temperature directly and allow thawing for 1-3 minutes.
• Add the appropriate volume of culture medium and centrifuge the cells at the standard settings. Optionally, cultivate the cells directly without centrifugation, but dilute the cryopreserved volume with at least a 50x volume of culture medium;
• For electroporation: Add 1-5 ug DNA/10e6 cells and place the suspension in an electroporation cuvette. Afterelectroporation at optimised settings, dilute with at least a 50x volume of culture medium at room temperature;