HaCaT-ras II-4 Cells

DMEM (Dulbecco's Modified Eagle Medium) is a highly versatile and widely utilized basal medium designed to support the growth of a diverse range of mammalian cells in biological research. It serves as an ideal medium for culturing primary fibroblasts, neurons, glial cells, HUVECs, smooth muscle cells, as well as popular cell lines like HeLa, 293, Cos-7, and PC-12.
What sets DMEM apart from other media is its unique composition. It contains an impressive fourfold increase in amino acid and vitamin concentration compared to the original Eagle's Minimal Essential Medium. Initially developed with low glucose (1 g/L) and sodium pyruvate, DMEM is frequently employed with higher glucose levels, either with or without sodium pyruvate. Notably, DMEM does not contain proteins, lipids, or growth factors, necessitating supplementation. To achieve optimal growth, a common approach is to supplement DMEM with 10% Fetal Bovine Serum (FBS). Additionally, DMEM employs a sodium bicarbonate buffer system (3.7 g/L), requiring a 5-10% CO2 environment to maintain a physiological pH.
Dulbecco's Modified Eagle Medium (DMEM) is highly regarded among defined media for cell and tissue culture, catering to the growth needs of various adherent cell phenotypes. It surpasses the original Eagle's Medium, developed in the 1950s for cultivating chicken cells, through the enhanced supplementary formulation known as Dulbecco's modification. This modification significantly elevates the content of select amino acids and vitamins up to fourfold compared to the original medium.
In the field of cell culture, DMEM plays a vital role as a growth medium for different cell types, including primary cells, stem cells, and transformed cells. Researchers also employ the modified version of DMEM for a wide array of research applications, such as drug discovery, tissue engineering, and the study of cell signaling pathways.
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 anhydrous
200,00


 
Iron (III) nitrate x 9H2O
0,10


 
Magnesium sulfate anhydrous
97,66


 
Potassium chloride
400,00


 
Sodium chloride
6.400,00


 
Sodium dihydrogen phosphateanhydrous
108,69


Other Components
D(+)-Glucose anhydrous
4.500,00


 
Sodium pyruvate
110,00


 
Phenol red
15,00



NaHCO3
1.500,00


Amino acids
L-Arginine x HCl
84,00


 
L-Cystine x 2HCl
62,58


 
L-Glutamine
584,00


 
Glycine
30,00


 
L-Histidine x HCl x H2O
42,00


 
L-Isoleucine
104,80


 
L-Leucine
104,80


 
L-Lysine x HCl
146,20


 
L-Methionine
30,00


 
L-Phenylalanine
66,00


 
L-Serine
42,00


 
L-Threonine
95,20


 
L-Tryptophan
16,00


 
L-Tyrosine x Na
103,79


 
L-Valine
93,60


Vitamins
D-Calcium pantothenate
4,00


 
Choline chloride
4,00


 
Folic acid
4,00


 
myo-Inositol
7,00


 
Nicotinamide
4,00


 
Pyridoxine x HCl
4,00


 
Riboflavin
0,40


 
Thiamine x HCl
4,00

Cytion’s Freeze Medium CM-1 is a state-of-the-art cryopreservation medium designed to ensure the highest level of cell viability and functionality post-thaw. This versatile medium is suitable for a broad spectrum of cell types, including both human and animal cells, making it an essential tool for diverse research applications. Formulated with a meticulously balanced combination of cryoprotectants and essential nutrients, Freeze Medium CM-1 minimizes ice crystal formation and cellular stress during the freezing process, thus preserving cellular integrity.
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.

Accutase Cell Dissociation Reagent
- 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) Solution: The Optimal Buffer for Your Biological Research
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