Marek: Freeze-down/Thaw

Overview

This short protocol describes how to freeze down and thaw (or bring up) mammalian cells (e.g. HeLa, KEK293, CHO, Jurkat T cells).

Materials

List reagents, supplies and equipment necessary to perform the protocol here.

• 10-50 ml centrifuge tube
• DMSO, Tissue culture (TC)-tested (e.g. Sigma, D2650)
• Cell culture medium (depends on cell line/cells)
• Trypsin-EDTA (e.g. from PAA, L11-004) - for adeherent cells
• Freeze-down medium
  • 6% DMSO
  • Cell culture medium (you can use 100% fetal calf serum (FCS) instead of cell culture medium)
• Basic TC centriguge (0 - 2500 rpm)
• Freeze-down box (this could be simply made from a small styropore box and cotton or from two styropore racks from 15-ml centrifuge tubes )

Procedure

1. Step 1
2. Step 2
   • Step 2 has some additional information that goes with it. i.e. Keep at 4°C.
3. Step 3
   1. Step 3 has multiple sub-steps within it.
   2. Enumerate each of those.

Notes

1. List troubleshooting tips here.
2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
3. Anecdotal observations that might be of use to others can also be posted here.

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Contact

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or instead, discuss this protocol.