100 mL $324.50
Store at 2°C – 8°C. SENSITIVE TO LIGHT.
EZ-CPZ™ is a superior GMP-manufactured medium for cryopreservation of human cells intended for clinical use or special research applications. It is a proprietary, chemically defined formulation of clinical grade components, cryoprotective agents and dimethylsulfoxide. EZ-CPZ™ is packaged in a convenient, ready-to use formulation, and is mixed 1:1 with concentrated cell suspensions so it does not require centrifugation.
Formulation and Packaging
EZ-CPZ™ is free of undefined or animal derived components, including blood products and proteins. EZ-CPZ™ has not demonstrated toxicity when injected into animals by various routes. Human and animal cells that are stored in EZ-CPZ™ and re-animated retain high viability and excellent growth and differentiation potential. EZ-CPZ™ has been developed for use with cells grown in serum-free media (such as INCELL’s ACE™ medium) where cryopreservation methods and components need to meet “animal component free” standards. When used in the 1:1 mix protocol, the final dimethylsulfoxide (DMSO) concentration is 5% v/v.
Use and Methods
EZ-CPZ™ is a high performance medium has been successfully used with many types of human and animal freshly isolated tissues and cells, in vitro cultured cells and established cell lines. These have been successfully cryopreserved in EZ-CPZ™ medium. Stored cells have been successfully reanimated with high recovery, viability, growth and differentiation capabilities.
Clinical Specimens and Early Passage Cultured Cells
This includes freshly isolated cells from multiple human solid tissues, blood cells, and tumor cells. Comparable results can be obtained from similar animal tissues. Cells initiated in culture from these sources with few or no subcultures, then stored in EZ-CPZ™, have a range of recovered viability from a low of 75% to 90% or greater in trypan blue dye exclusion assays. Cells from either adherent or suspension cultures are gently mixed 1:1 with EZ-CPZ™ at 1 to10 x106 cells/mL (depending on cell type), or other preferred cell density not to exceed 20% of the total volume of cryopreservation medium. Addition of serum or plasma is optional and is detrimental if the goal is to maintain a defined cell system. Tumor cells or blood cells freshly collected in media with or without their autologous (or a heterologous) plasma or serum with anticoagulant can also be mixed 1:1 with EZ-CPZ™. Alternatively, EZ-CPZ™ can be directly used as a standard 10% (v/v) DMSO cryopreservation medium to re-suspend cell concentrates or pellets. Cryovials or bags are placed in a controlled rate freezer container or in a “Mr. Freezy” with isopropanol at -80°C for at least 12 hours to cool down at about 1 degree per minute. They are then transferred to a –135°C freezer or liquid nitrogen freezer. Recovery of frozen cells is done rapidly by placing cryovials in a 37°C water bath for ~1 min until most of the medium has thawed. The cells are immediately placed in pre-warmed cell culture media. Cells are extremely fragile at this time and must be handled gently. Most cells recover from cryopreservation after they are grown for a few hours or overnight before the medium is changed. Although it is generally not necessary, some cells are centrifuged to remove the cryoprotectant media and then gently re-suspended in growth media.
INCELL’s proprietary E10 (anti-HHV-8) hybridoma, NCM460D™, NCM356D™, IBHK-4 and other standard cell lines have been successfully and repeatedly cryostored in EZ-CPZ™. It has also been used to cryostore numerous standard cell lines from many species and cell types, and grown in their recommended media. A range of examples include CHO, BHK, COS, 293, 3T3, U937, HT29 and HL60.
EZ-CPZ™ is manufactured using cGMP standards in an ISO Class 7 clean room and ISO Class 5 biosafety cabinet. All components are endotoxin-tested and filtered through a 0.22µm filter and checked for sterility (bacteria, fungi, and mycoplasma). Each lot of EZ-CPZ™ is tested for quality specifications and acceptance criteria. It is also performance-tested for cell viability and growth effects. A Certificate of Analysis assures that each lot meets specifications for acceptance.
- Acceptance Criteria
No microbial growth
No obvious particulate matter
Viability of E10 hybridoma cells ≥ 70% of starting viability
Storage of EZ-CPZ™
EZ-CPZ™ must be stored in a light-protective bag or other suitable container at 2° to 8°C. Do not freeze.
Master Files Applications Note
EZ-CPZ™ is in FDA Drug and Device Master Files but has not been tested by INCELL for any specific diagnostic or therapeutic use. To request use of a Master File call, fax, or email to [email protected]
Store at 2°C – 8°C. SENSITIVE TO LIGHT.