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Gibco™ PSC Cryopreservation Kit

Catalog No. A2644601
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A2644601 50 mL
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Catalog No. A2644601 Supplier Gibco™ Supplier No. A2644601
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Description

PSC Cryopreservation Kit

The PSC Cryopreservation Kit contains xeno-free PSC Cryopreservation Medium, which is a ready-to-use solution for the cryopreservation of early passage pluripotent stem cells (PSCs), and RevitaCell™ Supplement (100X), a chemically defined recovery supplement for use in the post-thaw culture media. When used in combination, these reagents help minimize loss of cell viability, maximize post-thaw recovery, and minimize unwanted differentiation of pluripotent stem cells. This kit can also be used to cryopreserve and recover peripheral blood mononuclear cells (PBMCs) to improve post-thaw cell viability and recovery.

  • Efficient recovery—routinely achieves 50% more viable cells than traditional methods of PSC cryopreservation
  • Consistent—chemically defined media maintains normal morphology, pluripotency, and karyotype of pluripotent stem cells over 10 passages
  • Reduced variability—optimized reagents minimize the causes of genetic instability of pluripotent stem cells

    Reliable
    Xeno-free, chemically defined formulations help ensure the proliferation capacity and long-term culture of PSCs and PBMCs in a stable state, thus helping to increase their usefulness in research and clinical research applications.

    Easy to use
    The PSC Cryopreservation Medium is provided as a ready-to-use, complete solution containing 10% DMSO for efficient cell addition. Separate additions of enhancing reagents are not required, helping save valuable preparation time.

    Versatile
    The kit is compatible with cells cultured in a variety of media systems including Essential 8™ medium, StemPro™ hESC SFM, and other commercially available formulations. Also, the kit is compatible with clump and single cell passaged PSCs.

    cGMP manufacturing and quality system
    The PSC Cryopreservation Kit is manufactured at a cGMP-compliant facility. The facility is also registered with the FDA as a medical device manufacturer and certified to ISO 13485 standards.

    • Specifications

    Classification Xeno-free for Cryomedium, Animal Origin-free for RevitaCell
    Product Type PSC Cryopreservation Kit
    Serum Level Serum-free
    With Additives DMSO (10%)
    Manufacturing Quality cGMP for medical devices, 21 CFR Part 820 and ISO 13485
    Product Line Gibco
    Quantity 50 mL
    Shipping Condition Dry Ice
    Content And Storage • 50 mL Cryopreservation Media
    • 5 mL RevitaCell Supplement (100X)

    Store at -5 to -20°C and protect from light.
    Culture Type Stem Cell (Human, iPS - Induced Pluripotent Stem, Embryonic)

    Frequently Asked Questions (FAQs)

    Can RevitaCell Supplement be used in addition to a traditional ROCK Inhibitor?

    No. Combination of RevitaCell Supplement with a traditional ROCK inhibitor will result in deleterious effects on PSCs.
    The RevitaCell Supplement in the PSC Cryopreservation Kit has a strong smell. Is this normal?

    Yes, it is normal that the RevitaCell Supplement has a strong smell.
    What is the difference between the PSC Cryopreservation Kit and the CTS PSC Cyropreservation Kit?

    The CTS PSC Cryopreservation Kit (Cat. No. A4239301) provides the same efficient recovery, consistency, and reduced variability as the PSC Cryopreservation Kit (Cat. No. A2644601). However, these two kits have different intended use statements, i.e., “For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. CAUTION: Not intended for direct administration into humans or animals” and “For Research Use Only”, respectively. Furthermore, the CTS PSC Cryopreservation Kit has full USP sterility testing info and a Drug Master file (DMF), whereas the sterility testing of the PSC Cryopreservation is via a modified USP method, and it does not have a DMF.
    Can I cryopreserve the iPSCs?

    You can cryopreserve iPSCs just as you would cryopreserve any pluripotent stem cells. Growth medium with 10% DMSO is recommended for freezing, or our PSC cryopreservation kit (Cat. No. A2644601) can be used
    Can the Cryopreservation Medium from PSC Cryopreservation Kit be aliquoted and frozen again?

    We have not tested this. However, the medium is stable when stored at 4 degrees C for up to 6 months. There are no components that R&D would be concerned about during a freeze thaw; however, this was not formally tested.
    What is the PSC Cryopreservation Kit?

    The PSC cryopreservation kit contains xeno-free PSC Cryopreservation Medium, which is a ready-to-use solution for the cryopreservation of early passage pluripotent stem cells (PSCs), and Gibco Revitacell Supplement (100X), a chemically defined recovery supplement for use in the post-thaw culture medium. When used in combination, these reagents help minimize loss of cell viability, maximize post-thaw recovery, and minimize unwanted differentiation of PSCs. This kit can also be used to cryopreserve and recover peripheral blood mononuclear cells (PBMCs) to improve post-thaw cell viability and recovery.
    Was the tissue cryopreserved prior to isolation of primary cells?

    No, the tissue was not cryopreserved.
    How do I establish a culture from cryopreserved cells?

    The procedure given below is a sample protocol for establishing cultures from the contents of one vial.

    1. Prepare a beaker of water at 37 degrees C.
    2. Remove a vial of cells from liquid nitrogen storage, taking care to protect hands and eyes.
    3. Loosen the cap on the vial 1/4 turn for 10 seconds to release any liquid nitrogen that may be trapped in the threads, then re-tighten the cap.
    4. Dip the lower half of the vial into the 37 degrees C water to thaw.
    5. When the contents of the vial have thawed, wipe the outside of the vial with disinfecting solution and move to a Class II, type A laminar flow culture hood.
    6. Open the vial and pipette the suspension up and down with a 1 mL pipette to disperse the cells.
    7. Remove 20 µL from the vial and dilute the cell suspension in 20 µL of trypan blue solution (for example: Gibco Trypan Blue, Cat. No. 15250-061).
    8. Use a hemacytometer to determine the number of viable cells per mL.
    9. Dilute the contents of the vial (1 mL) to the concentration recommended by the product instructions (for example 1.25 X 10E4 viable cells/mL for Gibco neonatal human epidermal keratinocytes ).
    10. Add 5 mL of cell suspension to each 25 cm2 culture flask or 15 mL of cell suspension to each 75 cm2 culture flask.
    11. Following inoculation, swirl the medium in the flasks to distribute the cells. Many cell types attach to culture surfaces quickly, and if the medium is not distributed immediately following inoculation, the cells may grow in uneven patterns.
    12. Incubate the cultures in a 37 degrees C, 5% CO2/95% air, humidified cell culture incubator. For best results, do not disturb the culture for at least 24 hours after the culture has been initiated.

    How I do thaw frozen cells?

    Please see our protocol here for thawing frozen cells (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/thawing-cells.html).
    How do I freeze my cells?

    Please see our protocol here for freezing cells (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/freezing-cells.html).

    For Research Use Only. Not for use in diagnostic procedures.