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Serum-Free Cell Freezing Medium

30-2600

ATCC® Serum-Free Cell Freezing Medium is a sterile, ready-to-use medium suitable for the cryopreservation of adherent and suspension cell cultures. This proprietary, serum-free formulation contains 10% DMSO and methylcellulose.

Cells cryopreserved using Serum-Free Freezing Medium show levels of viability and percent attachment (adherent cells) that are comparable to cells preserved in DMSO and FBS. Serum-Free Cell Freezing Medium can be used for both cells cultured in serum-supplemented growth medium as well as cells grown under serum-free conditions.
Product category
Reagents
Comments

Animal Component-Free, Protein-Free Cell Freezing Medium

Cells cryopreserved using Serum-Free Freezing Medium show levels of viability and percent attachment (adherent cells) that are comparable to cells preserved in DMSO and FBS. Serum-Free Cell Freezing Medium can be used for both cells cultured in serum-supplemented growth medium as well as cells grown under serum-free conditions.

Applications
Cell culture
Cell growth and viability
Cryopreservation
Volume
20 mL
Storage conditions
2°C to 8°C
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Documentation

ATCC determined that a biosafety level is not applicable to this material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to complete your own risk assessment and understand any potential hazards associated with the material per your organization’s policies and procedures and any other applicable regulations as enforced by your local or national agencies.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific applications
ATCC® Serum-Free Cell Freezing Medium is a sterile, ready-to-use medium suitable for the cryopreservation of adherent and suspension cell cultures.

Cells cryopreserved using Serum-Free Freezing Medium show levels of viability and percent attachment (adherent cells) that are comparable to cells preserved in DMSO and FBS. Serum-Free Cell Freezing Medium can be used for both cells cultured in serum-supplemented growth medium as well as cells grown under serum-free conditions.

Handling information

Handling procedure
Cryopreservation of Cells
Cryopreserve cells when cultures are actively growing. If growing adherent cells under serum-free conditions, we recommend the use of ATCC® 30-2101 Trypsin-EDTA Solution (1X) and ATCC® 30-2104 Soybean Trypsin Inhibitor (50X Concentrate) to detach the cells.
  1. Harvest the culture to prepare a cell suspension using your standard, cell-specific method.
  2. Centrifuge the cells at 125 x g for 5 to 10 minutes.
    1. Do not over-centrifuge cells as this may cause cell damage.
    2. After centrifugation, the cells should form a clean loose pellet.
  3. Aspirate medium and (neutralized) dissociation solution (used with adherent cells) and resuspend the cell pellet in 2 to 8 mL fresh, pre-warmed, complete growth medium.
  4. Count the cells. Centrifuge the cells again at 125 x g for 5 to 10 minutes.
  5. Take the Serum-Free Cell Freezing Medium from storage and swirl to mix. Decontaminate the vial by dipping in or spraying with 70% alcohol.
  6. Aspirate the medium and suspend the cell pellet in Serum-Free Cell Freezing Medium at a concentration 3 x 106 to 5 x 106 cells/mL. Aliquot 1 mL of the cell suspension to each labeled cryovial.
  7. Freeze the cells gradually at a rate of -1°C/min until the temperature reaches -70°C to -80°C. The cells should not be left at -80°C for more than 24 to 48 hours. Once at -80°C, frozen cryovials should be transferred to the vapor phase of liquid nitrogen for long-term storage.
Handling Procedure for Frozen Cells and Initiation of Cultures
  1. Warm the complete growth medium to 37°C prior to use with the cells.
  2. Thaw a vial of cells cryopreserved in Serum-Free Cell Freezing Medium by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 90 seconds).
  3. Remove the vial from the water bath before the contents are completely thawed, and decontaminate by dipping in or spraying with 70% ethanol. 
    All of the operations from this point on should be carried out under strict aseptic conditions.
  4. Transfer the vial’s contents plus 5 mL of complete growth medium to a 15 mL centrifuge tube. Use an additional 1 mL of medium to rinse the vial and transfer the liquid to the 15 mL tube. Add 4 mL of complete cell growth medium to bring the total volume to 10 mL.
  5. Spin the cells at 125 x g for 5 min. Aspirate the supernatant and resuspend the pellet in 2 mL of complete growth medium.
  6. Count the cells; adjust the volume so that the cells are plated at the appropriate seeding density.
Handling notes
This proprietary, serum-free formulation contains 10% DMSO and methylcellulose. Cells cryopreserved using Serum-Free Freezing Medium show levels of viability and percent attachment (adherent cells) that are comparable to cells preserved in DMSO and FBS. Serum-Free Cell Freezing Medium can be used for both cells cultured in serum-supplemented growth medium as well as cells grown under serum-free conditions.

Quality control specifications

Mycoplasma contamination
Not detected

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Frequently Asked Questions

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