HaCaT growing culture
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Culturing cells is crucial in biological research, as it allows scientists to study cellular behavior, growth, and development. Dulbecco's Modified Eagle Medium (DMEM) is a widely used basal medium that has proven effective in supporting the growth of a variety of mammalian cells, including primary fibroblasts, neurons, glial cells, HUVECs, and smooth muscle cells, as well as cell lines such as HeLa, 293, Cos-7, and PC-12.
What is DMEM and its Composition?
DMEM is a modified form of the original Eagle's Minimal Essential Medium, developed by Harry Eagle and published in 1959 in Science. The modification includes an increased concentration of amino acids and vitamins, making it a more nutritious medium for cell growth. DMEM uses a sodium bicarbonate buffer system, which helps maintain a physiological pH in a 5–10% CO2 environment.
Various DMEM formulations are available, including those with higher glucose levels, with or without sodium pyruvate, and L-glutamine/GlutaMAX supplementation. The standard DMEM formulation contains four mM L-glutamine, 4500 mg/L glucose, one mM sodium pyruvate, and 1500 mg/L sodium bicarbonate. It is important to note that DMEM commonly requires fetal bovine serum (FBS) supplementation for optimal growth of cells.
Choosing the Right DMEM for Your Experiment
In conclusion, Dulbecco's Modified Eagle Medium (DMEM) is a widely used basal medium that provides essential nutrients to support the growth of a variety of mammalian cells. With a range of formulations and options for customization, DMEM is an indispensable tool for cell culture experiments.
In biological research, the cryopreservation of mammalian cells is an invaluable tool. Successful preservation of cells is a top priority given that losing a cell line to contamination or improper storage conditions leads to lost time and money, ultimately delaying research results. Once the cells have been transferred from a cell growth medium to a freezing medium, the cells are typically frozen at a regulated rate and stored in liquid nitrogen vapor or at below -130°C in a mechanical deep freezer. The freeze medium CM-1 enables cryopreservation of cells at below -130°C (or in liquid nitrogen), essentially eliminating the need for an additional, costly ultralow freezer and eliminating time-consuming and demanding controlled rate freezing processes. Simply collect the cells, aspirate the growth medium, resuspend in CM-1, transfer to a cryovial, and store the vial at below -130 °C.
CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Trusted by hundreds of researchers
Our advanced cell freezing medium CM-1 is a market-leading product in Germany and Europe and is distinguished by numerous publications involving hundreds of different cell lines worldwide. We tested it with more than 1000 cell lines from our proprietary cell bank.
Optimized serum-free ingredients
CM-1 does contain serum products. Serum-containing cryopreservation mediums optimally protect the cells whilst being frozen and have the advantage of high recovery rates. As CM-1 has been tested with a multitude of cell lines, you can rest assured that your cells always recover well.
Contains FBS, DMSO, glucose, salts
Buffering capacity pH = 7.2 to 7.6
Applications & Validation
The cells preserved in our CM-1 freeze medium can be used for cell counting, viability and cryopreservation, cell culture, mammalian cell culture, gene expression analysis and genotyping, in vitro transcription, and polymerase chain reactions. Each batch's efficacy is evaluated using CHO-K1 cells. Each batch is tested for pH, osmolality, sterility, and endotoxins to ensure high quality.
- 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 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.