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Whole Tissue Cryoprotective Media (WTCM); DMSO Free
Cryopreservation of tissues especially when are big could be challenging. Moreover, the biggest obstacle is forming ice crystals. As a result, it causes volume expansion with the destruction of cell membranes. Whole Tissue Cryoprotective Media (WTCM); DMSO Free properly penetrates in the tissue and rapidly lowers the temperature inside the tissue. Under the DMSO Free condition, tissues must be frozen at the rate of 1°C/ min.
Description
Background
Cryopreservation of tissues, especially when they are large, could be challenging. Moreover, the biggest obstacle is forming ice crystals. Undeniably, it causes volume expansion with the destruction of cell membranes and holes in loose connective tissue. Whole Tissue Cryoprotective Media (WTCM); DMSO Free properly penetrates the tissue and rapidly lowers the temperature inside the tissue. Under the DMSO Free condition, tissues must be frozen at the rate of 1°C/ min.
The crucial Role of the Rapid freezing in whole Tissue Cryoprotection
Tissues should be frozen as rapidly as possible to promote the formation of amorphous ice (rather than crystal ice). With a low specific heat constant, liquid nitrogen, when it contacts warm tissues, boils and forms a vapor barrier which acts as an insulator. Consequently, the inner core of the tissue sample freezes slowly and unevenly. The PurMa™ WTCM has high thermal conductivity and therefore, has overcome the mentioned problem.
Our Cryoprotective strategy
PurMa Biologics has developed a DMSO-free whole-tissue cryoprotective media which is the outcome of many years of cutting-edge science. The cryoprotectant in our media is a complex of heavy chain sugars. This combination adequately freezes down the tissue with minimal ice formation and extracellular hydration.
Please read our protocol before using DMSO Free PurMa™ WTCM Cryoprotective Media as following the protocol is crucial for proper cryopreservation.
PurMa™ Whole Tissue Cryoprotective Media (WTCM)
- Ready-to-Use.
- It is DMSO-Free.
- PurMaTM WTCM provides much higher cell viability after short/long-term cryoprotection.
- PurMaTM WTCM Prevents the formation of crystals in extracellular space, consequently, preventing cellular dehydration and resulting in less exposure to toxins from the non-recovering cells.
- Preserves morphological appearance as a standard factor of cell recovery.
- Tested for pH, osmolality, sterility, and endotoxin.
- Each Lot of PurMaTM WTCM besides the pathogens and sterility evaluations goes through a survival test (by Trypan Blue method), viability & performance evaluation (by MTT Test), and the rate of recovery by Alamar Blue test.
- PurMaTM WTCM has been tested successfully for several commonly used tissue including rodent heart, brain, and kidney
Guideline
for PurMa Biologics guideline for cryoprotection, click here as well as in corresponding tab: PurMa™ Guideline for Cryoprotecting various cell lines
References
- Cryopreservation of whole ovine ovaries with pedicles as a model for human: parameters of perfusion with simultaneous saturations by cryoprotectants. Isachenko et al. Clin Lab. 2015;61(3-4):415-20. doi: 10.7754/clin.lab.2014.140919.
- Analysis of variations in cryomedium and methods of freezing on human bone marrow stem cells. O’Kane-Murphy et al.Prog Clin Biol Res. 1992;377:361-6.
| Parameter | Specification |
|---|---|
| Appearance | Red, clear liquid |
| pH | 7.2 ± 0.1 |
| Osmolality | 275-360 mOsm/L |
| Endotoxin | NMT< 2EU/mL |
| Mycoplasma | Negative |
| Suitability | Suitable for mammalian cell culture |
| Additive | Sodium pyruvate |
| Indicator | Phenol red |
| Mycoplasma Detection | Negative |
| Sterility Tested | Sterile filtered using 0.22 µm filter |
| Form | Liquid |
| Shipping Condition | Room temperature |
Learn more: What Is Cryoprotective Media?
Additional information
| Size | 1 x 10 ml, 1 x 100 ml |
|---|
Documents
Certificate of Analysis (COA): Available upon request for your specific order — email info@purmabiologics.com and we will send the current lot documentation.
Safety Data Sheet (SDS): Available upon request — email info@purmabiologics.com.
What our customers say about PurMa Biologics
Feedback from labs we supply, across our full catalogue.
We buy all of our cell culture media, fetal bovine sera, and lipid-depleted sera from PurMa. The quality of their reagents is absolutely top-notch, giving us reliable, repeatable results in our experiments. We highly recommend PurMa!
We have been working with PurMa biologics for the past several years, we buy cell culture related media like DMEM F12 and EMEM and Fetal Bovine Serum. We appreciate the high quality and reasonable price, and great customer service.
We purchase everything we need related to cell and tissue culture from PurMa, reagents like: DMEM, LL15, EMEM, FBS, trypsin, antibiotics mix100X and... Always great quality, fast processing and shipping. We are totally satisfied.
Very fast, efficient, and economic service. Great for new PIs like me.
it is reliable vendor!
We always buy our agarose from PurMa and have never been disappointed.
Frequently asked questions
Should I choose the DMSO or DMSO free formulation?
DMSO is the standard cryoprotectant and gives the best recovery for most lines. Choose DMSO free where DMSO would interfere with the application, for example in differentiation work or certain clinical and hybridoma protocols where residual DMSO is a problem.
What freezing rate should I use?
Approximately minus 1 degree C per minute to around minus 80, using a controlled rate cooler or an insulated freezing container, before transfer to liquid nitrogen. Faster cooling causes intracellular ice and slower cooling causes osmotic damage.
How should vials be stored long term?
In the vapour phase of liquid nitrogen, below minus 150 degrees C. Storage in a minus 80 freezer is acceptable only for weeks, not years, because viability declines steadily above minus 130.
How should I thaw cells frozen in Whole Tissue Cryoprotective Media (WTCM); DMSO Free?
Thaw rapidly in a 37 degree C bath until only a small ice crystal remains, then dilute promptly into pre warmed medium. DMSO is toxic to cells at room temperature, so the interval between thaw and dilution should be as short as possible.
How should the cryoprotective media itself be stored?
Store as stated on the Certificate of Analysis, protected from light. Do not refreeze after thawing the media, and use within the period stated for the lot.
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