Colo-824 Cells
General information
Description | The cells do not tolerate DMSO. Upon thawing, DMSO has to be removed immediately by centrifugation. |
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Organism | Human |
Tissue | Breast, mammary gland |
Disease | Squamous cell carcinoma |
Metastatic site | Pleural effusion |
Synonyms | COLO-824, COLO 824, Colo 824, COLO #824, COLO824, Colo824, Colorado 824 |
Characteristics
Age | 52 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Adherent/suspension |
Identifiers / Biosafety / Citation
Citation | Colo-824 (Cytion catalog number 300463) |
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Biosafety level | 1 |
Expression / Mutation
Protein expression | p53 negative |
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Tumorigenic | Yes, in nude mice |
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 |
Passaging solution | Accutase |
Doubling time | 3 days |
Subculturing | Gather the suspension cells in a 15 ml tube and gently wash the adherent cells with PBS lacking calcium and magnesium (use 3-5 ml for T25 flasks and 5-10 ml for T75 flasks). Apply Accutase (1-2 ml for T25 flasks, 2.5 ml for T75 flasks) ensuring full coverage of the cell layer. Allow the cells to incubate at room temperature for 10 minutes. Following incubation, combine and centrifuge both the suspension and adherent cells. After centrifugation, carefully resuspend the cell pellet and transfer the cell suspension into new flasks containing fresh medium. |
Split ratio | A ratio of 1:4 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
|
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: 10,12
D13S317: 12,13
D16S539: 11,12,13
D5S818: 12
D7S820: 8,11
TH01: 7,9
TPOX: 6,11
vWA: 16
D3S1358: 16.17
D21S11: 28
D18S51: 15.19
Penta E: 7
Penta D: 5,10
D8S1179: 12,14
FGA: 22
|
HLA alleles |
A*: 01:01:01, 34:02:01
B*: 13:02:01, 44:03:01
C*: 04:01:01, 06:02:01
DRB1*: 07:01:01, 15:03:01G
DQA1*: 01:02:01, 02:01:01
DQB1*: 02:02:01, 06:02:01
DPB1*: 04:01:01, 105:01:01
E: 01:01:01
|
Required products
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.
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate (2.0 g/L) and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
The unique composition of this RPMI formulation comprises 2.1 mM of stable Glutamine, 2.0 grams per liter of NaHCO3, and phenol red.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition
Components
mg/L
Inorganic Salts
Calcium nitrate x 4H2O
100,00
Magnesium sulfate anhydrous
48,83
Potassium chloride
400,00
Sodium chloride
5,950.00
di-Sodium hydrogen phosphate
800,49
Other Components
D(+)-Glucose anhydrous
2,000.00
Glutathione (red.)
1,00
Phenol red
5,00
NaHCO3
2,000.00
Amino Acids
L-Arginine x HCl
241,86
L-Asparagine x H2O
56,82
L-Aspartic acid
20,00
L-Cystine x 2HCl
65,19
L-Alanyl-L-Glutamine
447,00
L-Glutamic acid
20,00
Glycine
10,00
L-Histidine x HCl x H2O
20,27
L-Hydroxyproline
20,00
L-Isoleucine
50,00
L-Leucine
50,00
L-Lysine x HCl
40,00
L-Methionine
15,00
L-Phenylalanine
15,00
L-Proline
20,00
L-Serine
30,00
L-Threonine
20,00
L-Tryptophan
5,00
L-Tyrosine x 2Na
28,83
L-Valine
20,00
Vitamins
p-Aminobenzoic acid
1,00
D-(+)-Biotin
0,20
D-Calcium pantothenate
0,25
Choline chloride
3,00
Folic acid
1,00
myo-Inositol
35,00
Nicotinamide
1,00
Pyridoxine x HCl
1,00
Riboflavin
0,20
Thiamine x HCl
1,00
Vitamin B12
0.005
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate (2.0 g/L) and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
This RPMI 1640 medium contains 4.5 grams per liter of glucose.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition
Components
mg/L
Inorganic Salts
Calcium nitrate x 4H2O
100,00
Magnesium sulfate anhydrous
48,83
Potassium chloride
400,00
Sodium chloride
5450,00
di-Sodium hydrogen phosphate
800,49
Other Components
D(+)-Glucose anhydrous
4500,00
Glutathione (red.)
1,00
HEPES
2383,00
Phenol red
5,00
Sodium pyruvate
110,00
Amino Acids
L-Arginine x HCl
241,86
L-Asparagine x H2O
56,82
L-Aspartic acid
20,00
L-Cystine x 2HCl
65,19
L-Glutamine
300,00
L-Glutamic acid
20,00
Glycine
10,00
L-Histidine x HCl x H2O
20,27
L-Hydroxyproline
20,00
L-Isoleucine
50,00
L-Leucine
50,00
L-Lysine x HCl
40,00
L-Methionine
15,00
L-Phenylalanine
15,00
L-Proline
20,00
L-Serine
30,00
L-Threonine
20,00
L-Tryptophan
5,00
L-Tyrosine x 2Na
28,83
L-Valine
20,00
Vitamins
p-Aminobenzoic acid
1,00
D-(+)-Biotin
0,20
D-Calcium pantothenate
0,25
Choline chloride
3,00
Folic acid
1,00
myo-Inositol
35,00
Nicotinamide
1,00
Pyridoxine x HCl
1,00
Riboflavin
0,20
Thiamine x HCl
1,00
Vitamin B12
0,01
NaHCO3
1500,00
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