CLS-ACI-1 Cells
General information
Description | The CLS-ACI-1 cell line was established in 1998 from a solid mammary carcinoma, which was induced in a model organism through oral administration of 7,12-dimethylbenzo[a]anthracene (DMBA) at a dosage of 20 mg per kilogram body weight. DMBA is a well-known potent mutagen and carcinogen that is commonly used in experimental oncology for the induction of cancers, particularly in studies related to breast cancer. The establishment of the CLS-ACI-1 cell line from the tumor tissue allows for extensive in vitro exploration of breast cancer biology, particularly in understanding the mechanisms of carcinogenesis initiated by chemical agents like DMBA. In vitro studies using the CLS-ACI-1 cell line provide crucial insights into the cellular pathways and genetic alterations associated with mammary carcinomas. This cell line serves as a valuable tool for oncological research, including drug testing, resistance mechanisms, and cellular response to pharmacological agents. As a continuous cell line, CLS-ACI-1 offers a consistent and replicable model for studying the progression and treatment of breast cancer, facilitating the development of more effective therapeutic strategies against similar carcinomas induced by chemical agents in humans. |
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Organism | Rat |
Tissue | Breast |
Disease | Adenocarcinoma |
Synonyms | CLS-ACI-I |
Characteristics
Age | 3 months |
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Gender | Female |
Morphology | Epithelial-like |
Growth properties | Adherent/suspension |
Identifiers / Biosafety / Citation
Citation | CLS-ACI-1 (Cytion catalog number 500459) |
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Biosafety level | 1 |
Expression / Mutation
Oncogenes | Overexpression of Mycn gene. |
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Tumorigenic | Yes, in nude mice, ACI-rat |
Karyotype | Near triploid. 88.4% showing 51-69 chromosomes, 5% 38-50 chromosomes, 6.6% near tetraploid or higher ploidy level. |
Handling
Culture Medium | DMEM:Ham's F12, w: 3.1 g/L Glucose, w: 1.6 mM L-Glutamine, w: 15 mM HEPES, w: 1.0 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
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 1:3 to 1:5 is recommended |
Seeding density | 2 x 10^4 cells/cm^2 will yield in a confluent layer in about 6 to 7 days |
Fluid renewal | Every 3 to 5 days |
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) |
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 |
Rat_D1Wox31: 100
Rat_D2Wox37: 156
Rat_D19Wox11: 228
Rat_D10Wox8: 266,27
Rat_D4Wox7: 141,145
Rat_D2Wox27: 223
Rat_D5Rat33: 116,120,122
Rat_D10Wox11: 156,159
Rat_D1Wox23: 226,23
Rat_D12Wox1: 410
Rat_D6Wox2: 100,112,120
Rat_D8Wox7: 161,182
Rat_D6Cebr1: 239,241
SRY: x,x
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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.
This unique formulation combines Dulbecco's Modified Eagle Medium (DMEM) and Ham's F-12 (Ham's Nutrient Mixture F-12) in a precise 1:1 ratio. The addition of L-glutamine further enhances its composition.
DMEM, derived from Eagle's Minimal Essential Medium (EMEM), offers an increased concentration of amino acids and vitamins compared to its predecessor. In contrast, Ham's F-12 is based on Ham's F-10 medium, providing a complementary set of essential components.
To support optimal cell growth, it is common practice to supplement DMEM:Ham's F12 with FBS at a typical concentration of 5-10%. This addition is necessary as the medium lacks growth hormones, lipids, and proteins crucial for cellular development.
DMEM:Ham's F12 incorporates a pH buffer system and is often supplemented with phenol red, a pH indicator. Cultured cells in DMEM:Ham's F12, or any medium utilizing the bicarbonate buffer system, require a controlled CO2 environment of 5-10% to maintain appropriate pH levels. Phenol red enables monitoring of pH changes from 6.2 (yellow) to 8.2 (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 as well as 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
154,45
Iron(III)-nitrate x 9H2O
0,05
Iron(II)-sulfate x 7H2O
0,42
Potassium chloride
311,83
Copper(II)-sulfate x 5H2O
0.001
Magnesium chloride anhydrous
28,57
Magnesium sulfate
48,85
Sodium chloride
6,999.50
Sodium dihydrogen phosphate anhydrous
54,35
di-Sodium hydrogen phosphate
70,98
Zinc sulfate x 7H2O
0,43
Other Components
D(+)-Glucose anhydrous
3,151.00
HEPES
3,574.50
Hypoxanthine
2,04
Linoleic acid
0,04
DL-68-Lipoic acid
0.103
Sodium pyruvate
110,00
Phenol red
8,10
Putrescin x 2HCl
0.081
Thymidine
0,36
Amino Acids
L-Alanine
4,45
L-Arginine x HCl
147,35
L-Asparagine x H2O
7,50
L-Aspartic acid
6,65
L-Cystine x 2HCl
31,29
L-Cysteine x HCl x H2O
17,56
L-Glutamine
365,00
L-Glutamic acid
7,35
Glycine
18,75
L-Histidine x HCl x H2O
31,48
L-Isoleucine
54,37
L-Leucine
58,96
L-Lysine x HCl
91,37
L-Methionine
17,24
L-Phenylalanine
35,48
L-Proline
17,27
L-Serine
26,25
L-Threonine
53,55
L-Tryptophan
9,02
L-Tyrosine x 2Na x 2H2O
55,81
L-Valine
52,66
Vitamins
D-(+)-Biotine
0,004
D-Calcium pantothenate
2,12
Choline chloride
8,98
Folic acid
2,66
myo-Inositol
12,51
Nicotinamide
2,02
Pyridoxine x HCl
2,03
Riboflavin
0,22
Thiamine x HCl
2,17
Vitamine B12
0,68
NaHCO3
1,200.00
- 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.
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