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cell culture techniques
The European Collection of Authenticated Cell Cultures (ECACC) was established in 1984 as a cell
culture collection to service the research community and provide an International Depository Authority
recognised patent depository for Europe. Over the last three decades ECACC has expanded and diversified to become one of the premier collections of authenticated cell cultures in the world and this remains the core of ECACC’s business. The collections currently hold over 40,000 cell lines representing 45 different species, 50 tissue types, 300 HLA types, 450 monoclonal antibodies and at least 800 genetic disorders.
The development and maintenance of such a diverse collection has inevitably produced a high level of
specialist knowledge and this has enabled ECACC to position itself as a centre of expertise in all aspects
of cell culture. ECACC has developed a comprehensive range of cell culture services and diversified into
new product areas such as extracted from cell lines.
ECACC is the largest of four collections of clinically significant organisms, maintained by Public Health
England (PHE). The other collections are the National Collection of Type Cultures (NCTC), the National
Collection of Pathogenic Viruses (NCPV), and the National Collection of Pathogenic Fungi (NCPF).
Design and Equipment for the Cell Culture Laboratory 2.1 Laboratory Design
Perhaps one of the most undervalued aspects of tissue culture is the need
to design the facility to ensure that good quality material is produced in a
safe and efficient manner. Most tissue culture is undertaken in laboratories
that have been adapted for the purpose and in conditions that are not ideal.
However, as long as a few basic guidelines are adopted this should not
compromise the work.
Traditional methods of central nervous system culture
Cell and tissue culture techniques have been central to neuroscience research and the advancing knowledge of key mechanisms underpinning central nervous system (CNS) development, function and malfunction. These methods have permitted investigation of cellular level functions that would be infeasible to carry out routinely in vivo. Not only are animal studies often more time consuming and expensive, the complexity of the in vivo environment can make it difficult to isolate and control confounding variables in order to investigate specific cell and molecular features. In some cases inaccessibility of relevant tissue renders such studies completely impossible.
One of the earliest reports of in vitro CNS cell culture was carried out by Harrison, who observed neurite extension in fragments of frog medullary tube cultured using the hanging drop method, which involves the inversion of tissue-containing solution that has been placed on a cover slip to create a hanging drop [1]. The study was pivotal in representing one of the first examples of CNS tissue being grown ex vivo. By the mid-20th century, human CNS tissue culture had become more common and was often performed in either hanging drop or roller tube formats [2,3]. Studies carried out using roller tube methods involved growing neural tissue explants on rotating surfaces to allow alternating exposure to culture medium and gaseous phases.
In 1955, Cavanaugh became the first to report the application of monolayer culture using neurons derived from chick embryos and then, two decades later by the mid-1970s, monolayer CNS tissue culture had begun to gain popularity. The monolayer cell culture technique relied on dissociation of brain tissues using trypsin or collagenase to form single cell suspensions which were then grown on various glass or plastic substrates. These studies were used to identify morphological, molecular and electrophysiological features of neurons and other CNS relevant cell types in vitro (reviewed in).
Conventional monolayer culture methods have also been referred to as two-dimensional (2D) because they require cell adherence to a flat surface, to which culture medium is applied allowing access to nutrients that enable cell survival and proliferation. While healthy cells remain attached to the surface, necrotic cells become detached and are easily removed during media change. Monolayer tissue culture has since been widely adopted as a routine method for assaying cell activity. Although glass was initially the predominant culture substrate of choice, clear polystyrene has taken precedence due to its cost efficiency, ease of visual analysis, and ability to promote cell adherence without the use of coatings..
500 ml Basal medium with the growth factor supplement
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Description: Epithelial Cell Medium is a complete medium designed for the culture of epithelial cells. It was tested and optimized with epithelial cell growth and proliferation in vitro. It is formulated for use with 5% CO2 and 95% air in a humidified incubator.The medium consists of 500 ml of basal medium (containing essential and non-essential amino acids, vitamins, organic and inorganic compounds, hormones, growth factors, trace minerals), supplemented with epithelial cell growth supplement, antibiotics, and fetal bovine serum.Epithelial Cell Medium Supplement Kit includes:- 0.5 ML RI-Transferrin-Selenium (ITS)- 0.5 ML EGF- 5.0 ML L-Glutamine- 5.0 ML Antibiotic-Antimycotic Solution- 10.0 ML FBS*Before use, add Supplement Kit into 500ml basal medium.
Description: Pluronic® F-127 is a nonionic surfactant that is 100% active and relatively non-toxic to cells, and frequently used with dye AM esters to improve their water solubility.