Freezing down cells guards against unplanned mutations, senescence, and contamination. While unprotected cell freezing is ordinarily lethal, cell freezing with cryopreservation media helps protect cells for long-term viability as well as minimizes freezing injury for an intact cell supply on your schedule.
View protocols for cell freezing and thawing for more information.
The use of a freezing medium should contain a cryoprotective agent like dimethyl sulfoxide (DMSO) or glycerol. These agents reduce culture-medium freezing points and allow for slower cooling rates; greatly reducing the risk of ice crystal formation which can damage and kill untreated cells.
While each of the following Gibco freezing media are optimized for different cell types, they are all ready-to-use and contain 10% DMSO.
|Synth-a-Freeze Cryopreservation Medium||Recovery Cell Culture Freezing Medium||PSC Cryopreservation Kit|
|Tested cell types||Human keratinocytes, ESCs, MSCs, NSCs, other primary cell types||CHO-S, CHO-K1, HEK 293, Jurkat, NIH 3T3||Pluripotent stem cells, peripheral blood mononuclear cells|
|Not suitable for||Melanocytes||—||—|
|Chemical properties||Animal origin-free||Contains FBS||Xeno-free (Cryomedium)|
Animal origin-free (RevitaCell)
A complete, ready-to-use freezing medium, the Recovery medium formulation avoids messy mixing. Using an optimized version of the classic DMEM, serum, and DMSO, Recovery cell freezing medium is ready to go when you are. When treated with Recovery medium, cells experience improved recovery and viability post-thaw.
Method: For each of four different cell lines (i.e., CHO, MDCK, Swiss 3T3, HeLa), the same number of cells were frozen into each of seven different cryopreservation media. Upon thawing, each cell sample was diluted into the same volume of a growth medium and seeded in 12-well plates. The initial seeding densities were 2.3×104 cells/well for adherent cell lines, and 6.0×105 cells/well for non-adherent cell lines, based on cell counts determined at the cryopreservation step.
When the leading culture for a particular cell line had reached late log growth and none of the other cultures from that cell line had passed peak density, all cultures for that cell line were harvested and the data were analyzed. Thus, different cell lines were analyzed on different days: CHO, day 6; MDCK, day 6; Swiss 3T3, day 5; and HeLa, day 6. Cell recovery of replicate cultures from different cryopreservation media were normalized to Recovery Cell Culture Freezing Medium and plotted (average normalized recovery±standard error).
Result: For cryopreservation of both adherent and suspension cell lines, Recovery freezing medium results in an average of 25% higher cell viability compared to other cell freezing media:
Flyer: Recovery freezing medium cell viability
Formulated without antibiotics, antimycotics, hormones, growth factors, serum or proteins, Synth-a-Freeze medium is defined and ready-to-use. Synth-a-Freeze medium is buffered with HEPES and sodium bicarbonate and designed for use with primary mammalian cell types including human keratinocytes, embryonic stem cells, neural stem cells, and mesenchymal stem cells.
NOTE: Synth-a-Freeze medium is not appropriate for melanocytes.
The PSC Cryopreservation Kit contains ready-to-use, xeno-free cryopreservation medium as well as a bottle of RevitaCell Supplement (100X). Together, these reagents minimize cell viability loss and unwanted cell differentiation while maximizing post-thaw recovery. Applications include pluripotent stem cells cultured in media systems, such as Essential 8 and StemPro hESC SFM, as well as other commercially available formulations. In addition, this kit can also be used to cryopreserve and recover peripheral blood mononuclear cells (PBMCs).
Containing Rho-associated protein kinase (ROCK) inhibitors and antioxidant properties, the RevitaCell supplement typically recovers pluripotent stem cells at 40,000 viable cells/cm2, and significantly reduces the average post-thaw recovery time of most cryopreservation solutions. In addition to pluripotent stem cells, RevitaCell supplements can also be utilized with primary cells including neurons, human epidermal keratinocytes, and human corneal epithelial cells.
Gibco cell culture products are manufactured in facilities compliant with current good manufacturing practices (GMP) and adhere to a robust quality management system, helping to ensure the consistency, reliability, and high quality you can rely on.
The 1 L Gibco bottle uses 38% less source material (32% less material for the 500 mL bottle) than comparable media bottles on the market today. This helps reduce the consumption of non-renewable resources, helping shrink your footprint.
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