Colo-680N Cells
















General information
Description | COLO-680N is a human esophageal squamous cell carcinoma cell line derived from the tumor biopsy of a 58-year-old woman in 1985. This derivation was unique as it involved passage through a nude mouse, a method used to enhance tumor cell growth and adaptation in vitro by leveraging the immunodeficient nature of the mouse. This process potentially selects for more aggressive and clinically relevant cancer cells, making COLO-680N particularly valuable for studying the intricate biology of esophageal squamous cell carcinoma, a major subtype of esophageal cancer. |
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Organism | Human |
Tissue | Esophagus |
Disease | Squamous cell carcinoma |
Applications | BMP-6 expression can be used as a co-indicator for prognosis in esophageal squamous‐cell carcinoma. In-vitro platform for long-term cultivation of Cryptosporidium parvum |
Synonyms | COLO 680N, COLO #680N, COLO680N, Colorado 680N |
Characteristics
Age | 57 years |
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Gender | Female |
Ethnicity | African |
Morphology | Epithelial-like |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | COLO-680N (Cytion catalog number 300464) |
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Biosafety level | 1 |
Expression / Mutation
Protein expression | BMP-6 |
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Handling
Culture Medium | RPMI 1640, w: 2.0 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 | 60 hours |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:4 is recommended |
Seeding density | 2 x 10^4 cells/cm^2 will yield in a confluent layer in about 4 to 5 days |
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 | As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. |
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: 11,12
D13S317: 13
D16S539: 11,12
D5S818: 11
D7S820: 10,12
TH01: 8
TPOX: 6
vWA: 17,18
D3S1358: 15
D21S11: 27
D18S51: 19
Penta E: 7,8
Penta D: 12
D8S1179: 14,15
FGA: 18.2
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HLA alleles |
A*: '02:01:01, '30:02:01
B*: '15:16:01, '57:01:01
C*: '06:02:01, '14:02:01
DRB1*: '07:01:01, '11:01:02
DQA1*: '01:01:02, '02:01:01
DQB1*: '03:03:02, '05:01:01
DPB1*: '01:01:02, '04:01:01
E: '01:01:01
|
Required products
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
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 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.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +8°C, protected from light.
Once opened, store at 4°C and use within 6–8 weeks.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Amino Acids
Glycine
10.00
L-Alanyl-L-Glutamine
434.40
L-Arginine
200.00
L-Asparagine H2O
56.82
L-Aspartic Acid
20.00
L-Cystine 2HCl
65.20
L-Glutamic Acid
20.00
L-Histidine HCl H2O
20.27
L-Hydroxy-L-Proline
20.00
L-Isoleucine
50.00
L-Leucine
50.00
L-Lysine 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 2Na 2H2O
28.83
L-Valine
20.00
Vitamins
p-Amino Benzoic Acid
1.00
D-Biotin
0.20
Choline Chloride
3.00
D-Calcium Pantothenate
0.25
Folic Acid
1.00
myo-Inositol
35.00
Nicotinamide
1.00
Pyridoxine HCl
1.00
Riboflavin
0.20
Thiamine HCl
1.00
Vitamin B12
0.005
Inorganic Salts
Ca(NO3)2 4H2O
100.00
KCl
400.00
MgSO4 7H2O
100.00
NaCl
6000.00
NaHCO3
2000.00
Na2HPO4
800.00
Other Components
D-Glucose
2000.00
L-Glutathione Reduced
1.00
Phenol Red Sodium Salt
5.30
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a widely used buffer solution in biological and chemical research. It plays a crucial role in maintaining the pH balance and osmolarity during various experimental procedures, including tissue processing and cell culture. Our PBS solution is meticulously formulated with high-purity ingredients to ensure stability and reliability in every experiment. The osmolarity and ion concentrations of our PBS closely mimic 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:
8000 mg/L Sodium chloride (NaCl)
200 mg/L Potassium chloride (KCl)
1150 mg/L Sodium phosphate dibasic anhydrous (Na2HPO4)
200 mg/L Potassium phosphate monobasic anhydrous (KH2PO4)
This composition ensures an optimal pH and ionic balance, suitable for a wide range of biological applications.
Applications of Our PBS Solution
Our PBS solution is ideal for various applications in biological research. Its isotonic and non-toxic properties make it suitable for substance dilution and cell container rinsing. PBS solutions containing EDTA are effective for disengaging attached and clumped cells. However, divalent metals such as zinc should not be added to PBS, as this can cause precipitation. In such cases, Good's buffers are recommended. Additionally, our PBS solution is an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, including SARS-CoV-2.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +25°C, protected from light.
Once opened, store at 2°C to 25°C and use within 24 months.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Salts
Potassium chloride
200
Potassium phosphate monobasic anhydrous
200
Sodium chloride
8000
Sodium phosphate dibasic anhydrous
1150