CCRF-CEM Cells
General information
Description | CCRF-CEM cells are a type of human T lymphoblasts commonly used in immuno-oncology and immunology research. These cells were isolated from the peripheral blood of a 4-year-old female Caucasian with acute lymphoblastic leukemia (ALL). CCRF-CEM grow in suspension and can reach high cell density when cultured in spinner flasks. Karyotype analysis of CCRF-CEM cells showed a modal number of 47 chromosomes, ranging from 41 to 95. They show no consistent loss or gain of specific chromosomes and no marker chromosomes. However, 28% of cells with 45 chromosomes showed C- and 53% of all cells had an extra D, and 35% had an additional F. CCRF-CEM cells are tumorigenic and can cause tumours in Syrian hamsters. These cells express CD3, CD5, CD7, and CD4 genes and antigens. Additionally, isoenzyme analysis showed ADA, 1; ES-D, 1; G6PD, B; GLO-I, 1; PEP-D, 1; PGD, C; PGM1, 1; PGM3, 0. These cells are reported to be free of virus particles as determined by electron microscopy. A study has shown that the combination of resveratrol and prednisolone induced apoptosis in CCRF-CEM cells in a time- and dose-dependent manner. The combination treatment showed synergistic effects on the overexpression of BAx and the downregulation of BCL2. |
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Organism | Human |
Tissue | Peripheral blood |
Disease | Leukemia |
Synonyms | CCRF/CEM, CCRFCEM, CCRF.CEM, CCRF CEM, CCRF, CEM, CEM-CCRF, CEM-CCRF (CAMR), CCRF/CEM/0, CEM/0, CEM-0, CCRF-CEM/S, GM03671, GM03671C |
Characteristics
Age | 4 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Polymorph cells, big nuclei, formation of microvilli |
Cell type | T lymphoblast |
Growth properties | Suspension |
Identifiers / Biosafety / Citation
Citation | CCRF-CEM (Cytion catalog number 300147) |
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Biosafety level | 1 |
Expression / Mutation
Protein expression | p53 negative |
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Antigen expression | CD3 B (37%), CD4 (50%), CD5 (95%), CD7 (77%) |
Isoenzymes | G6PD, B |
Tumorigenic | Yes, in nude mice |
Viruses | EBV negative |
Reverse transcriptase | negative |
Ploidy status | Aneuploid |
Handling
Culture Medium | RPMI 1640, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) |
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Medium supplements | Supplement the medium with 10% FBS |
Doubling time | 24 hours |
Subculturing | Maintain cultures by periodically adding or replacing the medium. Initiate cultures with a density of 2 x 10^5 cells/ml and keep the cell concentration within the range of 1 x 10^5 to 1 x 10^6 cells/ml for optimal growth. |
Seeding density | Start new cultures at 1 x 10^5 cells/ml |
Fluid renewal | Every 3 days |
Freezing recovery | Allow the cells to recover from the freezing process for at least 48 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,x
CSF1PO: 10,13
D13S317: 11
D16S539: 10,13
D5S818: 12,13
D7S820: 9,13
TH01: 6,7
TPOX: 8
vWA: 17,19
D3S1358: 14,15
D21S11: 30,34.2
D18S51: 13,18
Penta E: 5,14
Penta D: 10,11
D8S1179: 12,13
FGA: 23,24
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HLA alleles |
A*: 01:01:01, 07:01:02
B*: 08:01:01, 16:01:02
C*: 03:04:01, 07:01:01
DRB1*: 03:01:01, 07:01:01
DQA1*: 02:01:01, 05:01:01
DQB1*: 02:01:01, 02:02:01
DPB1*: 04:01:01, 13:xx
|
Required products
Key Features of Freeze Medium CM-ACF
Serum-Free Formulation: Eliminates the variability and risks associated with serum, providing a defined and controlled environment for a wide array of cell types, including primary cells, stem cells, and established cell lines.
Broad Compatibility: Effective for a wide array of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-ACF is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Using CM-ACF for Freezing Cells
To use CM-ACF for freezing both adherent and suspension cells, follow these steps:
Preparation:
For adherent cells: Wash and dissociate them from the culture substrate.
For suspension cells: Proceed directly to the next step.
Cell Counting: Ensure cells are at the proper concentration.
Centrifugation: Pellet the cells and resuspend them in CM-ACF freeze medium.
Cryovial Transfer: Transfer the resuspended cells into cryovials.
Freezing Process: Use a slow-freezing method before transferring the cells to long-term storage.
Freezing Methods
Method
Description
Steps
❄️ Manual Freezing
A step-by-step method involving gradual temperature reduction to ensure cell viability.
1. Place cells in freeze medium in a 4°C freezer for 40 minutes. 2. Transfer to a -80°C freezer for 24 hours. 3. Store cells in liquid nitrogen for long-term preservation.
🧊 Using Mr. Frosty
A convenient device that allows for controlled freezing rates without electrical power.
1. Prepare cells in cryovials with freeze medium. 2. Place cryovials in Mr. Frosty container. 3. Store at -80°C for 24 hours before transferring to liquid nitrogen.
🧬 Controlled-Rate Freezer
A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.
1. Program the device to gradually decrease the temperature. 2. Place prepared cells in the freezer. 3. After the freezing cycle, transfer cells to liquid nitrogen.
Long-Term Storage
Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.
Ingredients
Contains DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6
Cytion’s Freeze Medium CM-ACF offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.
Key features of Freeze Medium CM-1 include:
Broad Compatibility: Effective for a wide range of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Using CM-1 for Freezing Cells
To use CM-1 for freezing both adherent and suspension cells, follow these steps:
For adherent cells, wash and dissociate them from the culture substrate. For suspension cells, proceed directly to the next step.
Count the cells to ensure they are at the proper concentration.
Centrifuge the cells to pellet them, then resuspend in CM-1 freeze medium.
Transfer the resuspended cells into cryovials.
Use a slow-freezing method before transferring the cells to long-term storage.
🥶 Method
🔍 Description
💡 Steps
❄️
Manual Freezing
A step-by-step method involving gradual temperature reduction to ensure cell viability.
1️⃣ Place cells in freeze medium in a 4°C freezer for 40 minutes.
2️⃣ Transfer to a -80°C freezer for 24 hours.
3️⃣ Store cells in liquid nitrogen for long-term preservation.
🧊
Using Mr. Frosty
A convenient device that allows for controlled freezing rates without electrical power.
1️⃣ Prepare cells in cryovials with freeze medium.
2️⃣ Place cryovials in Mr. Frosty container.
3️⃣ Store at -80°C for 24 hours before transferring to liquid nitrogen.
🧬
Controlled-Rate Freezer
A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.
1️⃣ Program the device to gradually decrease the temperature.
2️⃣ Place prepared cells in the freezer.
3️⃣ After the freezing cycle, transfer cells to liquid nitrogen.
Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.
Ingredients
Contains FBS, DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6
Cytion’s Freeze Medium CM-1 offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.
Phosphate-buffered saline (PBS) is a versatile buffer solution used in many biological and chemical applications, as well as tissue processing. Our PBS solution is formulated with high-quality ingredients to ensure a constant pH during experiments. The osmolarity and ion concentrations of our PBS solution are matched to those of the human body, making it isotonic and non-toxic to most cells.
Composition of our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains 137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, and 2 mM KH2PO4. We have chosen this composition based on CSHL protocols and Molecular cloning by Sambrook, which are well-established standards in the research community.
Applications of our PBS Solution
Our PBS solution is ideal for a wide range of applications in biological research. Its isotonic and non-toxic properties make it perfect for substance dilution and cell container rinsing. Our PBS solution with EDTA can also be used to disengage attached and clumped cells. However, it is important to note that divalent metals such as zinc cannot be added to PBS as this may result in precipitation. In such cases, Good's buffers are recommended. Moreover, our PBS solution has been shown to be an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, such as SARS-CoV-2.
Storage of our PBS Solution
Our PBS solution can be stored at room temperature, making it easy to use and access.
To sum up
In summary, our PBS solution is an essential component in many biological and chemical experiments. Its isotonic and non-toxic properties make it suitable for numerous applications, from cell culture to viral transport medium. By choosing our high-quality PBS solution, researchers can optimize their experiments and ensure accurate and reliable results.
Composition
Components
mg/L
Inorganic Salts
Potassium chloride
200,00
Potassium dihydrogen phosphate
200,00
Sodium chloride
8,000.00
di-Sodium hydrogen phosphate anhydrous
1,150.00