The application of insect cell culture for heterologous protein expression has progressively increased over the last several decades. The continued development of new cell culture technology is essential for the future growth and application of insect cell and baculovirus biotechnology. Pdf insect cell culture and biotechnology researchgate. Animal cell culture classical media insect cell culture. The essential elements for tissue culture mass production are insect cell lines that can support viral replication, a cell medium that can support cell culture, and suitable bioreactors for large. Compare insect cell lines from leading suppliers on biocompare. The development of the cell lines antheraea eucalypti cell lines the cultures from which these four lines developed were set up in 1960 and 1961. Insect cell culture an overview sciencedirect topics. Graces insect medium, unsupplemented 500 ml 11595030 graces insect medium, supplemented 500 ml 11605094 sf900 ii sfm 500 ml 10902096. Insect cell culture technology has become an important part of modern biotechnology. Browse sigmaaldrich s culture plate inserts to find products in corning transwell cell culture plate insertsa and accessories, corning netwells, corning snapwells, millicell inserts. A 100 ml spinner containing 50 ml of cell culture is at a density of 2 x 10 6 cellsml. Insect cell cultures are widely used in studies on insect cell. Expression systems is committed to supporting our customers through this challenging time.
Insect cell culture as a tool in plant virus research. Vaccines, antibiotics and other essential medicines have always constituted basic necessities for our society. This research service brings out the emerging trends in mammalian cell culture systems on a global scale, through a detailed description of. An important factor underscoring this popularity of insect cell expression is the innate ability of insect cells to produce relatively large quantities of posttranslationally modified eukaryoticproteins in a relatively short period of time. The use of insect cell lines as production hosts is an emerging technology for the production of bio pharmaceuticals. This paper describes techniques for the development of continuous cell lines from insect embryos and other tissues. Cell culture cell culture is one of the major tools used in cellular and molecular biology, providing excellent model systems for studying the normal physiology and biochemistry of cells e.
Find the right classical media formulation for your mammalian cell culture. All the powdered media and prepared liquid culture media should be stored at 28c. Efficient production and purification of recombinant. Mature late log or stationary phase cell culture 1, 25cm 2 tissue culture flasks 2, cell culture medium, 1, 2 and 5ml sterile pipets, sterile 200 l pipet tips, trypsin diluent divalent cationfree pbs, vmf trypsin 0. Cell culture basics handbook thermo fisher scientific. Insect cell culture media hyclone sfx insect cell culture media designed to support the growth of multiple insect cell lines and production of a variety of recombinant proteins. Insect viruses provide an effective control method against certain pests of oil palm plantations in the tropical countries. Insect cell culture if youre keen to establish your own culture of insect cells, our spodoptera frugiperda sf cells lines are the perfect place to start. We have thousands of liters of cell culture media esf 921 and esf af in stock and ready to ship both domestically and internationally. Methods for the development of cell lines from insects. This product is useful as a broad spectrum antibacterial and antifungal agent. Purifying histagged proteins from insect and mammalian cells. About your presenter kamal has over thirty years of academic experience in both research and biotechnology educational program development plus an additional focus on training at universities. Bioreactor conditions were chosen to maximize cell growth rate and all conditions and.
Uncontaminated cell cultures uncontaminated cell cultures do not appear to present a significant hazard as even direct dermal inoculation may result in only local inflammation. In the earliest attempts to grow insect tissues in vitro, the main interest was the study of the mechanisms of mitosis and meiosis 37. At this point, the culture will need to be disaggregated usually with proteolytic enzymes like trypsin into individual cells and subcultured split, passaged, or transferred. This manual is included with sf9, sf21, and high five insect cells and provides general information.
Remove 25 ml of medium and cells, replace with 25 ml of fresh complete medium for a final density of 1 x 10 6 cellsml. If your protein of interest is a secreted protein, it is best to use a serum or proteinfree insect cell culture for the production run, as described elsewhere murhammer, 2007. The study of basic cell biology, diseases and cancer, the development and testing of new therapeutics, and the production of new drugs relies on using the techniques introduced in this experiment. Proteolysis is a problem of the bevs system due to its lytic nature, and can, therefore, be a critical issue in insect cell bioprocessing. By using an animal componentfree medium, the possible threat of adventitious agent contamination. High level recombinant protein production in insect cell. Seed cultures were grown in spinner flasks, and the bioreactor was inoculated at a cell density of 2 x 105 cellsml. Confluency confluency is a marker for when to subculture your cells.
Many animal componentfree formulations have been developed for other cell culture platforms, such as cho and ns0. It has many promising areas of application like study of insect cell physiology and baculovirus biology, production of insect pathogenic viruses as insecticides for agricultural pest control and most importantly production of recombinant proteins. Insect cell culture oxford expression technologies. One culture set up in august 1960 contained four whole ovaries from diapausing pupae, whereas the cultures set up in october 1960, january 1961, and march 1961 contained cells from four. Production of entomopathogenic viruses in insect cell cultures. Various insect and mammalian cell culture media to verify that the magnehis niparticles are compatible with purification of histagged proteins from commonly used insect and mammalian cell culture media, purified histagged thermostable luciferase 5g was added to various culture media in the presence or absence of 10% fetal bovine serum. Insect cell culture is a common choice for heterologous protein expression. There are currently more than 100 insect cell lines available for recombinant protein production with lines derived from bombyx mori, mamestra brassicae, spodoptera frugiperda, trichoplusia ni, and drosophila melanogaster being of particular interest. The insect cell culture media is phosphate buffer based. Inspite of above recommended storage condition, certain. Pdf the continued development of new cell culture technology is essential for the future growth and application of insect cell and baculovirus. Endotoxin content nmt 5euml storage and shelf life. Insect cell culture and biotechnology springerlink. Key words insect cell lines, tissue culture, cell culture.
Expression systems insect cell culture media are designed for maximum virus and protein expression. Specialized media, transfection reagents, and vectors have been developed in response to recent advances in insect cell culture and molecular biology methods. Cell culture hyclone sfx insect cell culture media designed to support the growth of multiple insect cell lines and production of a variety of recombinant proteins. The culture of insect cells in the absence of serum is reaching maturity, and promising serum substitutes hydrolysates, new growth and productionenhancing factors are being evaluated. Cell culture is a vital technology used in life science research and in biotechnology laboratories.
Your use of this pdf, the bioone complete website, and all posted and associated content indicates. Following this first passage, the culture is generally referred to as a cell line. Jan 29, 2016 high level recombinant protein production in insect cell culture kamal rashid, ph. Culture plate inserts cell culture supplies sigmaaldrich. A high degree of purity is achievable with an easy purification process, particularly when recombinant protein is secreted into the cell culture medium.
The use of cell lines for academic research and for commercial applications is currently dominated by two cell lines. Jan 16, 2008 the continued development of new cell culture technology is essential for the future growth and application of insect cell and baculovirus biotechnology. Factors affecting the sialylation of fc fusion protein in. An essential of the baculovirus expression system, sf insect cells are an excellent choice for recombinant virus production and amplification, protein expression, and virus titration. A series of four cho cell culture batches,referred to as batches14, were conducted using a 2l autoclavable glassbioreactor. Insect cell culture for industrial production of recombinant. For largescale applications, the baculovirus expression vector system bevs is particularly advantageous. Insect cell culture media is more acidic than mammalian media ph 6. Mammalian cells, being extremely sensitive to growth conditions such as shear, require equipment and processes that. High level recombinant protein production in insect cell culture kamal rashid, ph. The basic reason for the failure was the lack of a suitable cell culture medium for culturing insect cells. Gibco insect media has been formulated for maximum growth and protein yields. Lynn1 usdaars, insect biocontrol laboratory, barcwest, building 011a, room 214, beltsville, maryland 207052350 summary the success of insect cell culture is demonstrated by reports of over 500 established cell lines. High level recombinant protein production in insect cell culture.
Insect cell culture gas mixer gas blender equipment. It is essential reading for all those with research interests in neuroscience, including developmental biologists, neurologists, nerve cell physiologists and neuropharmaoologists. Methods include how to obtain aseptic primary tissues, how to dissociate the tissue, and then how to maintain the primary culture until cells grow consistently. Being eukaryotic, insect cells allow correct folding and posttranslation modification, and purified protein can be used therapeutically. How many cells do you recommend to use for seeding a flask at 50% confluence for insect cells. The need of these goods, greatly increased over the last several decades, leads the modern science towards new reliable and efficient drug production methods. Home expression systems insect cell culture media and.
If the cell culture continues to grow and it is passaged at least ten times, it is considered a continuously replicating cell line. To initiate cell culture immediately upon receipt, see page 10. Mammalian cell culture biocon has established a largescale, multiproduct, cell culture facility which is the core technology in the manufacture of several important biologicals such as monoclonal antibodies and erythropoietin. Guide to baculovirus expression vector systems bevs and. For largescale production or basic research, insect cells are able to express large quantities of protein with complex posttranslational modifications. Continuous ocr in any incubator or workstation get continuous, noninvasive oxygen consumption rate ocr measurements for days to weeks from inside any incubatorworkstation that fits your cell culture plates. Maintain the insect cell culture density range between 1 x 10 6 1 x 10 7 cellsml by splitting and diluting the cell culture with fresh sf900 ii media. Pdf insect cell culture and applications to research and. The insect cell culture based protein expression system is inexpensive and easy to scale up. A confluent monolayer is an adherent cell culture dish, plate or flask in which the cells have formed a single layer over the entire surface area available for growth.
The sf9, sf21, and high fivez cell lines are suitable for use in expressing recombinant proteins with baculovirus and other insect expression systems e. The resipher series of cell culture monitors are the. Pdf on jan 1, 2010, spiros n agathos and others published insect cell culture find, read and cite all the research you need on researchgate. Growth and maintenance of insect cell lines user guide. It is a testament to their scientific insights that these media still form the basis of most mammalian cell culture research being performed today. These investigations required only simple media in which the. The neuron in tissue culture is a comprehensive reference source dealing exclusively with the tissue culture of neurons. Els article, insect cell culture by just m vlak and monique m van oers. Count the cells in a sample volume using a hemocytometer and light microscopy to calculate the culture cell. Guide to baculovirus expression vector systems bevs and insect cell culture techniques. This serumfree, proteinfree insect cell culture medium provides unmatched cell growth and expression for a wide range of insect cells including sf9, sf21, tni high fivetm and drosophila s2. The formulations created by cell culture pioneers like eagle, dulbecco, ham, and others can be found below. Additionally, in case of baculovirus constructs, protein yields can vary with virus titer, expression time after infection, cell culture medium, etc. Factors affecting the sialylation of fcfusion protein in recombinant cho cell culture gyun min lee, department of biological sciences, kaist, 335 gwahakro, yuseonggu, daejeon 305701, republic of korea over the past decade, product titers from recombinant cho cells have increased significantly.