Toxicity, membrane binding and uptake of the sclerotinia. Since then, more than 600 insect cell lines have been. We recommend sf9 or sf21 cells for transfection, purification, and amplification of recombinant virus. Aphidsymbiotic bacteria cultured in insect cell lines. These cell lines were generated from the parental cell line. Growth and maintenance of insect cell lines the wolfson centre. Points to consider on the characterization of cell lines. Established insect cell lines used for the production of recombinant proteins grow to higher densities than mammalian cells, thus smaller culture volumes are sufficient. Pdf insect cell culture and applications to research and. These cell lines have been used in diverse fields of research as described in the other chapters of this book.
Their sf9 cell line comes from the fall armyworm and is highly effective as a rapid growth. Atcc was entrusted with its first cell line in 1962 and has consistently attained the highest standards and used the most reliable procedures to verify every cell line since. Introduction to insect cell line strains spodoptera frugiperda sf9 and sf21 insect cells are cell lines commonly used for expression of recombinant proteins using baculovirus systems. Pdf ecdysone action on insect cell lines macro mark. The fingerprinting pattern is a stable characteristic of the cell line. Lynn summary this chapter lists the known cell lines from lepidoptera, largely based on previous compilations of insect cell lines published by w. All handling of insect cell lines should be carried out under sterile conditions, as you would for mammalian cell culture. Welcome high level recombinant protein production in insect cell culture webinar series 2. The sf9, sf21, and high fivez cell lines are suitable for use in expressing recombinant proteins with baculovirus and other insect.
While established procedures that can be used for developing new. Insect cell lines can grow over longterm passaging, which. The success of insect cell culture is demonstrated by reports of over 500 established cell lines. Insect cell lines manual the wolfson centre for applied structural. Insect cells are biohazard level 1, but are susceptible to bacterial infection if sterile technique is poor. Passagingsubculturing refers to diluting cells back to a density that maintains log phase growth and maximum viability. All the insect cell lines that we own have been adapted to serumfree media. Cell lines derived from embryos of the tobacco hornworm,manduca sexta l. Expression systems offers insect cell lines adapted to serumfree suspension culture in either esf 921 or esf af. Your use of this pdf, the bioone complete website, and all posted and. 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. Read this article to learn about the types, nomenclature, selection and maintenance of cell lines. The high five cell line btitn5b14 isolated from the cabbage looper, trichoplusia ni is an insect cell line widely used for baculovirusmediated recombinant protein expression. The national cancer institute nci seeks licensing partners for a novel modified insect cell line, sf9et, that can quickly and efficiently determine baculovirus titers during the.
Insect sf9 cells vigene has entered an exclusive cgmp partnership with virovek to use viroveks advanced bactoaav. Despite its widespread application in industry and academic laboratories, the genomic background of this cell line. The sf9, sf21, and high fivez cell lines are suitable for use in expressing recombinant proteins with baculovirus and other insect expression systems e. High level recombinant protein production in insect cell.
Toxicity, membrane binding and uptake of the sclerotinia sclerotiorum agglutinin ssa in different insect cell lines. In other words, once the primary culture is subcultured, it becomes a cell line. In the area of pathology, an important use of insect cell lines is in the investigation of virus cell interactions. Cell lines derived from primary cultures have a limited life span i. Mimic insect cells are qualified using the following criteria. Mimic cells lines are suitable for transfection and expression of. Development of a new animal componentfree, proteinfree. Insect cell culture an overview sciencedirect topics. 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. The field has grown to the extent that now over 500 insect. The official designation is given for each cell line. We suggest that the two bacteria persist in aphids because competition between them is limited by differences in tropism for insect tissues or cell. These studies have led to the application of cell lines. Identification of insect cell lines from 8 lepidopteran.
This paper describes techniques for the development of continuous cell lines from insect embryos and other tissues. Presented at cell culture engineering iv, engineering foundation conferences, san diego, ca 712 march 1994 abstract we2, cell. Development of a new animal componentfree, proteinfree medium for sf21 and sf9 insect cell lines z. Insect cell lines in suspension, cultivated in roller. This manual is included with sf9, sf21, and high five insect cells and provides general information about the. The term cell line refers to the propagation of culture after the first subculture. Cell line normal cells usually divide only a limited number of times before losing their ability to proliferate, which is a genetically determined event known as senescence. Using the advent of new techniques and instruments in molecular biology as well as the advancement in biotechnology, the indigenous cell lines. The relative advantages of different tissues embryos, ovaries, etc. Although insect cell cultures are less demanding than mammalian cells. Insect cell lines can be utilized in many areas of biological research, including physiology, toxicology, and pathology. Similarly, most of the commercially available insect cell lines grow well in monolayer or suspension culture. In cell lines infected with bacteria derived from aphids harboring both t type and u type, the u type persisted, while the t type was lost. The use of cell lines for academic research and for commercial applications is currently dominated by two cell lines.
Uncontaminated cell cultures uncontaminated cell cultures do not appear to present a significant hazard as even direct. Design of an efficient medium for insect cell growth and recombinant. This paper describes methods that are effective for maintaining various insect cell lines. For largescale production or basic research, insect cells are able to express large quantities of protein with complex posttranslational modifications. Growth and maintenance of insect cell lines user guide 9. Transcriptome analyses of insect cells to facilitate. A great number of cell lines from hemipteran, lepidopteran, and dipteran insects have been developed and characterized. As the field has advanced from methods development to a standard procedure, so has the diversity of scientists using the technique. Pdf in vitro infection of wolbachia in insect cell lines. Caple sigmaaldrich biotechnology, 3050 spruce street, saint louis, mo 63103 usa abstract the baculovirus expression vector system1 bevs, when used with insect cell lines.
Their sf9 cell line comes from the fall armyworm and is highly effective as a rapid growth medium. The insect cell culture based protein expression system is inexpensive and easy to scale up. Profacgens technical reference guide for recombinant protein expression in insect cells using the baculovirus system presents one of our current technical strategies, and can be used as a reference for insect cell. Being eukaryotic, insect cells allow correct folding and posttranslation modification, and purified protein can be used therapeutically. In addition, both cell lines achieved good growth in bioreactor batch culture and had a prolonged stationary phase of 34 d in ypr medium compared to insect. Pdf mariner transposable element in insect cell lines. Methods for the development of cell lines from insects.
The atcc cell biology collection is one of the largest bioresources in the world, and offers a complex array of human, animal, insect, fish and stem cell lines. Cell line characterization dna barcode assay for cells that are to be used in gmp biomanufacturing processes, confirmation of cell identity in a master cell bank mcb is required1,2. Growth and maintenance of insect cell lines user guide pub. In this chapter, i will provide a brief overview of how new cell lines can be established and, once obtained, how they should be handled and characterized. High level recombinant protein production in insect cell culture 1. Although wolbachia has not been cultured outside its host cells, it can be cocultured with insect cell lines in vitro, and this technique would facilitate studies on wolbachia. Cell culture basics handbook thermo fisher scientific. One such company was protein sciences whose recombinant dna platform is based on insect cells. Cell line a cell line is a permanently established cell culture that will proliferate indefinitely given appropriate fresh medium and space a cell culture developed from a single cell and therefore consisting of cells with a uniform genetic makeup there is presence of several cell. 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.
The development and various other aspects of primary culture are described above. New insect sf9et cell line for determining baculovirus. Dna amplification fingerprinting daf with arbitrarily selected primers was used to obtain dna fingerprint profiles to distinguish among 8 lepidopteran insect cell lines. Pdf insect cell culture and biotechnology researchgate. The cultivation of insect cell lines in suspension in roller bottles provides a way of obtaining large amounts of cells in a short period of time. Insect cell cultures are now commonly used in insect physiology, developmental biology, pathology, and molecular biology. The availability of preadapted cells saves significant time and expense. A high degree of purity is achievable with an easy purification process, particularly when recombinant protein is secreted into the cell. Methods cell handling techniques handle insect cell lines under sterile conditions in a laminar flow hood.
Development and characterization of insect cell lines. Sf9 cells, a clonal isolate of spodoptera frugiperda sf21 cells iplbsf21ae, are commonly used in insect cell culture for recombinant protein production using baculovirus. Sf9 cells are regular in size, easy to manipulate, and form good monolayers for plaque assays. Leading coronavirus vaccine development using cells of.