Dynamic gas mixtures with adjustable O2 and CO2 concentration in cell cultures study
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MCQ Gas Blender 100 Series High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface www.mcqinst.com Gas Blender 100 Series Application Note 1/4 Dynamic gas mixtures applications Working with static gas mixtures with fixed or adjustable amount of CO2 represents now just the base of the cell culture applications. Since the last two decades until today many experiments have been successfully conducted with gas mixtures dynamically obtained and many more demands for instruments capable of controlling both O2 and CO2 relative amounts in the culture atmosphere. Fine control over the gas mixture allow those experiments, in which an induced hypoxic, hyperoxic or hypercapnic state is required, to be performed efficiently. Introduction Starting from the pioneering studies made in the early 1900’s, the cell culture process has been developed and refined to become one of the most important and widely used experimental method for the scientific research. The growth of cells is a complex process that requires appropriate and controlled conditions (such as pressure, temperature, and system pH) to be achieved. A crucial role is carried out by the gas mixture fluxed over the culture. The instrument for the gas mixtures management must secure high precision outcomes over the whole experiement duration; high accuracy, great stability and high repeatability are required. On the other hand it’s also important to work with an instrument capable of creating gas mixtures dynamically adjustable since variation of the conditions for a particular cell type can result in gene/protein/enzyme expression changes. For all these applications we propose the use MCQ Gas Blender 100 Series, an instrument specifically made to work with 3 components gas mixtures that offers a quick and easy way to efficiently manage the experimental condition of your cell culture. Common experimental setup One of crucial aspect of the grown of cells in vitro is the complete control over the working parameters during the entire process. Even if the cell culture applications may vary widely from each other, growth medium represent one the most important of these parameter. The medium used is usually buffered with sodium bicarbonate/carbonic acid and for a proper cell grown the pH value must be strictly maintained constant. The amount of CO2 in the culture atmosphere directly affects the medium pH, therefore the control over the gas mixture in which cells are grown becomes another fundamental process parameter. Due to the low content of CO2 (<0,04%), the air isn’t a valid option for cell culture. In most of the common applications the cells are grown in an atmosphere of 5-10% CO2, inside special CO2 incubators. The basic CO2 incubators make use of gas mixture cylinders (pre-mixed by the supplier) in which the amount of CO2 in the flow cannot be changed or adjusted by the user. Newer and more advanced incubators include a CO2 sensor for a feedback regulation of the the amount of CO2 inside the chambers but are less common and more expansive then the basic ones. MCQ Gas Blender 100 Series Application: Dynamic gas mixtures with adjustable O2 and CO2 concentration in cell cultures study Application Note

MCQ Gas Blender 100 Series High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface www.mcqinst.com Gas Blender 100 Series Application Note 2/4 [3,4], the gene expression [5] and for enzyme studies [6], while the hyperoxic condition is primary induced over the cells for the lung injuries study [7]. A hypercapnic mixture is prepared varying CO2 concentration instead of O2. Basically a hypercapnic mixture contains 21% oxygen and an increased amount of CO2 compared with air. Studies conducted over cell cultures in hypercapnic condition have been useful for the collection...

MCQ Gas Blender 100 Series High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface www.mcqinst.com Gas Blender 100 Series Application Note 3/4 have been chosen for simplicity). The gas cylinders are connected to the instrument through 6 mm diameter tubes and a check valve is installed along each line as backflow prevention device. Each gas media is connected and controlled by one of the 3 channels of the Gas Blender 100. Another 6 mm tube finally connects the instrument to the working system in which the cells are grown. The image below shows an example of working...

MCQ Gas Blender 100 Series High Performance Gas flow Dilutor & Gas flow Mixing System with User Interface www.mcqinst.com Gas Blender 100 Series Application Note 4/4 References [1] J. G. Murphy, T. W. Smith and J. D. Marsh, Calcium flux measurements during hypoxia in cultured heart cells – J. Mol. Cell. Cardiol. 19, 3 (1987) 271-279. [2] I. Ginis, D. V. Faller, Hypoxia affects tumor cell invasiveness in vitro: the role of hypoxia-activated ligand HAL1/13 (Ku86 autoantigen) – Cancer Lett. 154, 2 (2000) 163-174. [3] M. Weinmann, P. Marini, V. Jendrossek, A. Betsch, B. Goecke, W. Budach and C....