G361 Cells




















General information
Description | G361 is a human melanoma cell line derived from a metastatic site in the skin of an adult patient. This cell line exhibits melanin production, which is a characteristic feature of melanocytes and melanoma cells. G361 cells are known for their epithelial-like morphology and are extensively used in research focused on skin cancer, particularly melanoma. The cells are valuable for studying the biology and pathogenesis of melanoma, including cell proliferation, migration, and invasion. Additionally, they serve as a useful model for drug screening and for understanding the mechanisms of resistance to chemotherapy in melanoma. The G361 cell line has been used to explore the genetic and molecular underpinnings of melanoma. It has been instrumental in studies investigating the role of various oncogenes and tumor suppressor genes in cancer progression. For instance, research utilizing G361 cells has contributed to insights into the MAPK/ERK pathway, which is often dysregulated in melanoma. These cells are also commonly used in assays to evaluate the efficacy of new therapeutic agents, making them crucial for translational research and the development of targeted treatments for melanoma. |
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Organism | Human |
Tissue | Skin |
Disease | Melanoma |
Synonyms | G-361, G361-mel, G361mel |
Characteristics
Age | 31 years |
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Gender | Male |
Ethnicity | European |
Morphology | Epithelial |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | G361 (Cytion catalog number 302157) |
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Biosafety level | 1 |
Expression / Mutation
Isoenzymes | G6PD, B |
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Products | Melanin |
Handling
Culture Medium | McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
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 | 1:2 to 1:4 |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) |
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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Benötigte Produkte
One prominent application of McCoy's 5A Medium is its utilization in the culturing of human colon carcinoma cell lines. Specifically, it has been employed in the study of the leucine-rich repeat-containing G-protein-coupled receptor (LGR5) and its role in the metastasis of colon cancer. This medium has been effectively employed in the cultivation of several colon carcinoma cell lines, including HCT116, RKO, FET, CBS, HCT116b, and TENN, enabling researchers to delve deeper into the mechanisms underlying colon cancer metastasis.
In addition to its application in cancer research, McCoy's 5A Medium has proven to be indispensable in the study of osteoblasts. Researchers investigating the ion reactivity of calcium-deficient hydroxyapatite in standard cell culture media have utilized this medium to culture osteoblasts. This application has facilitated a better understanding of the interactions between osteoblasts and calcium-deficient hydroxyapatite, contributing to advancements in the field of bone research.
Notably, McCoy's 5A Medium was meticulously formulated by modifying the amino acids found in Basal Medium Eagle to provide optimal support for liver tumor cells. This enriched formulation enables its suitability for a diverse range of established cell lines, as well as primary cells, further enhancing its versatility and applicability in various research settings.
Moreover, McCoy's 5A Medium extends its biochemical and physiological effects beyond liver tumor cells. It has been successfully employed to support growth in primary cultures of bone marrow, skin, gingiva, kidney, omentum, adrenal, lung, spleen, rat embryo, and other cell types. This wide range of applications attests to the broad utility of McCoy's 5A Medium in supporting the growth and maintenance of various cell types for comprehensive biological research.
Formulation
This McCoy's 5A medium (modified) contains 3.0 g/L of Glucose, stable Glutamine, 2.0 mM of Sodium pyruvate, and 2.2 g/L of NaHCO3.
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 chloride x 2H2O
132,46
Magnesium sulfate x 7H2O
200,00
Potassium chloride
400,00
Sodium chloride
6,460.00
Sodium dihydrogen phosphate x H2O
580,00
Other Components
D(+)-Galactose anhydrous
3,000.00
Glutathione (red.)
0,50
Bacto-Peptone
600,00
Phenol red
10,00
Amino Acids
L-Alanine
13,36
L-Arginine x HCl
42,10
L-Asparagine x H2O
45,00
L-Aspartic acid
19,97
L-Cysteine
24,24
L-Glutamine stable
326,61
L-Glutamic acid
22,10
Glycine
7,50
L-Histidine x HCl x H2O
20,76
L-Hydroxyproline
19,70
L-Isoleucine
39,36
L-Leucine
39,36
L-Lysine x HCl
36,50
L-Methionine
14,90
L-Phenylalanine
16,50
L-Proline
17,30
L-Serine
26,30
L-Threonine
17,90
L-Tryptophan
3,10
L-Tyrosine
18,10
L-Valine
17,60
Vitamins
p-Aminobenzoic acid
1,00
Ascorbic acid
0,50
D(+)-Biotin
0,20
D-Calcium pantothenate
0,20
Choline chloride
5,00
- 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.