RCC-GS Cells








General information
Description | Established from the kidney clear cell, pT3b, No, M1/ GIII (brain metastasis) carcinoma of a 56-years-old-male, 1999. |
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Organism | Human |
Tissue | Kidney |
Disease | Clear cell renal cell carcinoma, pT3b, No, M1/ GIII (brain metastasis) |
Synonyms | KTCTL-185, KTCTL185, RCCGS |
Characteristics
Age | 56 years |
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Gender | Male |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | RCC-GS (Cytion catalog number 300241) |
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Biosafety level | 1 |
Expression / Mutation
Surface antigens | Cytokeratine positive 8,18,19, vimentin positive |
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Protein expression | IL8 |
Tumorigenic | Yes, in nude mice |
Mutational profile | IL8 RS1126647 3-UTR SNP T>T |
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 | A ratio of 1:2 to 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 48 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: 10,11
D13S317: 8,14
D16S539: 9,12
D5S818: 11,12
D7S820: 8,10
TH01: 9
TPOX: 8
vWA: 16,18
D3S1358: 16
D21S11: 31
D18S51: 14
Penta E: 8,10
Penta D: 11
D8S1179: 8,11
FGA: 24
PEZ6: HROG36
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Required products
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
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