{"id":8625,"date":"2020-07-28T17:01:00","date_gmt":"2020-07-28T17:01:00","guid":{"rendered":"https://www.stirlingultracold.com/de/?p=8625"},"modified":"2021-07-31T19:58:19","modified_gmt":"2021-07-31T19:58:19","slug":"freezing-cells","status":"publish","type":"post","link":"https://www.stirlingultracold.com/de/freezing-cells/","title":{"rendered":"Process for Freezing and Thawing Cells"},"content":{"rendered":"\n\n\n\n

Requirements for Freezing and Thawing Cells 

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Established cell lines are a valuable resource for research labs. However, even the best-kept cell lines in a continuous culture can be susceptible to genetic drift, senescence, and microbial contamination. Therefore, the cells must be frozen and preserved for long-term storage. 

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Below are some guidelines that are essential for freezing cells. As with all cell culture procedures, it is best to follow the instructions provided with your cell line for optimal results. 

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Thawing frozen cells can be a complicated procedure in which it is essential to be both quick and precise in order to guarantee that most of the cells will survive. For the best possible outcome, it is recommended that you closely abide by the instructions given with your cells and other components. For best results, follow these guidelines:

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Materials Required for Thawing Cells 

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Step-by-step Process for Thawing Frozen Cells

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    \n
  1. Start by removing the cryovial that contains the frozen cells from liquid nitrogen storage or ultracold freezer.
    2. \n
  2. Place cell vial in a water bath at 37° C and gently swirl, until only a small amount of ice remains. Alternatively, use an automated cell thawing instrument such as the BioLife ThawSTAR® CFT1.5 or CFT2 by inserting the frozen cryogenic vial into the device. Retrieve it when the vial is raised at the end of the thaw cycle and proceed. 
    3. \n
  3. Keep in mind that it is important to decrease exposure to room temperature when you remove frozen cells from storage. If cells are not going to be used immediately, place the cells on dry ice, or in a liquid nitrogen container. Cells in 1.8 mL and 2.0 mL cryovials can be transported from frozen storage to the thawing device inside the BioLife ThawSTAR® CFT Transporter while frozen on dry ice. Alternatively, cryovials containing cells can be transferred frozen at <-150℃ inside the BioT™ LN2 Transporter, which safely contains liquid nitrogen within an absorbent/baffle pad.
    4. \n
  4. Move the vial into a laminar flow hood.
    5. \n
  5. Wipe the exterior of the vial with 70% ethanol or isopropanol.
    6. \n
  6. Transfer the preferred amount of pre-warmed complete growth medium appropriate for your cell line into the thawed cells. Thawed cells frozen in cryovials are generally pipetted out of the small cryovial into a centrifuge tube containing a pre-warmed growth medium.
    7. \n
  7. Centrifuge the cell suspension at 200 x g for around 5-10 minutes at room temperature. The speed and duration of the centrifugation will change based on the cell types.
    8. \n
  8. Once the centrifugation is complete, check the clarity of the supernatant as well as the visibility of a complete pellet. 
    9. \n
  9. Without disturbing the pellet, gradually pour the supernatant. 
    10. \n
  10. Remove the supernatant with a pipette and leave a small amount of medium to ensure the cell pellet is not disrupted.
    11. \n
  11. Carefully tap the tube to resuspend the cell pellet.
    12. \n
  12. Move them into their appropriate cell culture vessel as well as the cell culture environment suggested.
    13. \n
  13. Now the cells are ready to be used in downstream applications.
    14. \n
","protected":false},"excerpt":{"rendered":"

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