Introduction an important part of adherent cell culture media is serum, which aids in both cell attachment and cell growth. Hek293 cell culture and transfection hek293 cell line. Hek 293 cells can be cryopreserved in conditioned growth medium supplemented with 10% vv dmso at 1 x 10 6 cellsml. Intermediate filament characterization was carried out using rtpcr to determine the expression of keratin no. Canonical wnt signaling in endothelial cells ecs is required for vascularization of the central nervous system cns and for formation and maintenance of barrier properties unique to cns vasculature. Hek 293 hek, hek 293, hek293, hek293 human embryo kidney transformed with sheared human ad5 dna. The cells were obtained from a single, apparently healthy, legally aborted fetus under dutch law. This resource enables novel and more informed studies with 293 cells, and we will distribute the sequenced cell lines to this effect. Typically, these experiments involve transfecting in a gene or combination of genes of interest, and then analyzing the expressed protein. One of the most commonly used neuralorigin cells is the neuro2a cell line. The hek cell line has been extensively used as an expression tool for recombinant proteins since it was generated over 25 years ago. Evaluation of hek293 cell growth and adenovirus productivity in hyclone cdm4hek293 medium human embryonic kidney 293 hek293 cells are widely used in production of recombinant viruses and proteins because of their stable growth, high transfection. The kidney cells were forced to take up bits of dna using a technique invented in 1973 that used a calcium solution.
Freestyle 293 expression medium is an animal origin free, chemically defined, proteinfree medium specifically developed to support the growth and transfection of 293f cells in suspension without adaptation. Preferential transformation of human neuronal cells by human adenoviruses and the origin of hek 293 cells. The glosensor camp hek293 cell line was maintained for up to 20 passages on 10cm culture dishes in dmem growth medium containing 50. Hek 293 cells are very easy to grow and transfect very readily and have been widelyused in cell biology research for many years. Having a higher expression level or a higher protein yield is generally desirable.
They are also known, more informally, as hek cells. For the last 25 years, hek 293 or, in short, 293 cells have been a staple both for. Hek 293 f cells, whose parental cell line hek 293 has been used by researchers for decades, are a suitable production cell line for rhfviii and. Apr 10, 2002 these may be the cell type of origin of 293 and other ad transformed cell lines. Furthermore, they have the ability to release urodilatin, the structural analog to anp.
Hek293 is listed in the worlds largest and most authoritative dictionary database of abbreviations and acronyms the free dictionary. General information about the human embryonic kidney 293 hek293 cell line including information about hek293 origin, expression, and cytogenetics. Here, we evaluated the potential of transient transfection in producing rfviii in serumfree suspension hek 293 cell cultures and investigated the effects of different dna concentration 0. Fectopro transfection reagent for bioproduction polyplus. These human distal tubule cells contain specific mrna for the anpurodilatin receptor npragca, whereas the receptors for other members of the natriuretic family, gcb and gcc, were.
Recombinant protein expression for structural biology in. Hek 293293t cells are derived from human embryonic kidney transformed with adenovirus 5 dna hek cells are easy to culture and transfect, and contain the sv40 large tantigen, which allows for the episomal replication of plasmids containing the sv40 origin of replication notes hek cells are loosely adherent, and do not require digestion. Genome dynamics of the human embryonic kidney 293 lineage in. A hek 293 derived cell line had been engineered harboring a single target locus with open reading frames encoding the livingcell reporter proteins lacz and egfp, the latter translationally. Several descendants of the original hek293 cell line established by graham and coworkers in 1977 graham et al. The widespread use of this cell line is due to its transfectability by the various techniques, including calcium phosphate method. Hek293 cell lines human embryonic kidney 293 cells hek 293 are a popular and easytotransfect option in cell biology and biotechnology. A combination of immunostaining, immunoblot, and microarray analysis showed that 293 cells express the neurofilament nf subunits nfl, nfm, nfh, and ainternexin as well as many other proteins. Hek293 cell line origins, cytogenetics, and expression. Animal cells and cell lines, such as hek293 cells, are commonly cultured at 37c. The synergistic effect of hek 293 cells grown in serumfree. Hek 293 cells have a complex karyotype, exhibiting two or more copies of each chromosome and with a modal chromosome number of 64. As shown on table 1, hek293 cells but not mesangial cells expressed keratin no.
As a consequence, hek 293 cells should not be used as an in vitro model of typical kidney cells. Hek293 is a cell line derived from human embryonic kidney cells grown in tissue culture. Can be used to isolate transformation defective hostrange mutants of ad5 and for titrating human adenoviruses. These cells are often used to express recombinant proteins. Hek 293 cell, the human embryonic kidney 293 cells was originally derived from human embryonic kidney cells grown in tissue culture. Transient transfection of serumfree suspension hek 293. Transfection efficiencies generally approach 100 percent, and the resulting cultures can be used to produce viral vectors, vaccines, and recombinant proteins. Cloning and sequencing of the cellularviral junctions from the human adenovirus type 5 transformed 293 cell line. Halo custom edition maps, tags, tutorials and assets for the halo ce game halo custom edition halo utilities. For the last 25 years, hek 293 or, in short, 293 cells have been a staple both for virologists studying human adenoviruses ads and for anyone who needed an easily transfectable cell line.
Human embryonic kidney 293 cells, also often referred to as hek 293, hek293, 293 cells. Enhancing protein expression in hek293 cells by lowering. The synergistic effectof hek 293 cells grown in serumfree medium with the corningcellbindsurface helen l. That only a small fraction of cells in hek cultures are susceptible to ad transformation is consistent with the extreme difficulty in establishing the original 293 cell line and other ad transformed hek cells.
These cells have been derived from the hek 293 cell line, are adapted to grow in suspension cultures, reaching high densities using serumfree media such as freestyle 293 expression medium. What is the difference between hek293 and hek293t cell lines. Hek 293293t cells are derived from human embryonic kidney transformed with adenovirus 5 dna hek cells are easy to culture and transfect, and contain the sv40 large tantigen, which allows for the episomal replication of plasmids containing the sv40 origin of replication notes. These cells have been derived from the hek 293 cell line, are adapted to grow in suspension cultures, reaching high densities using serumfree media such as. The e1a adenovirus gene is expressed in these cells and participates in transactivation of some viral promoters, allowing these cells to produce very. Their ease of growth and transfection makes hek293 cells a common cell culture in academic. Hek 293 cells display a variety of features of the late distal tubule. Human embryonic kidney 293 hek293, hek293, or hek cells are one of the most common cell lines used for research.
The human embryonic kidney cell line hek293 is known to express several neuronal. Cells were maintained at 37 c in a 5% co 2 atmosphere and passed by trypsinization every 34 days to maintain subconfluence. Evaluation of hek293 cell growth and adenovirus productivity in hyclone cdm4hek293 medium human embryonic kidney 293 hek293 cells are widely used in production of recombinant viruses and proteins because of their stable growth, high transfection level, and ability to produce high viral titers. The synergistic effect of hek 293 cells grown in serum. Here we use wholegenome resequencing of six 293 cell lines to study the dynamics of this aneuploid genome in.
The 293 cell line was derived by transformation of primary cultures of human embryonic kidney hek cells with sheared adenovirus ad5 dna. Page 1 of 2 material safety data sheet for cell cultures biosafety level 1 1. Here we use wholegenome resequencing of six 293 cell lines to. Pdf preferential transformation of human neuronal cells by human. Apr 17, 2015 the cells, called hek 293 cells that stands for human embryonic kidney were taken from an aborted fetus in the 1970s in the netherlands. While 293 freestyle cells are a derivative of a 293 wildtype strain selected for good growth performance and high expression levels zhang et al.
Material safety data sheet for cell cultures biosafety level 1. Hek293 cells are useful for many transfection experiments, particularly the propagation of adenoviralbased and retroviralbased vectors. Cell viability is the number living cells relative to the total number of cells. Bits of chopped up dna from the adenovirus, a virus that causes a pretty severe cold.
Hek293 cells were isolated from primary human embryonic kidney tissue. The approach uses transient cotransfection of a human embryonic kidney hek cell line e. Human embryonic kidney 293 cells, also often referred to as hek 293, 293 cells, or less precisely as hek cells are a specific cell line originally derived from human embryonic kidney cells grown in tissue culture. Our enewsletter is a great way to stay up to date on the most recent news and products from cell biolabs. Utility of human embryonic kidney cell line hek293 for rapid. The scattered twelve tribes of hek293 hek293 the hek293 line is the original line established from a primary embryonic human kidney and transformed with sheared human adenovirus type 5 dna. Hek 293 cells are straightforward to grow in culture and to transfect. Although of epithelial origin, its biochemical machinery is capable of carrying out most of the posttranslational folding and processing required to generate functional, mature protein from a wide spectrum of. The popular human embryonic kidney hek 293 cell line and its derivatives have been used for decades in both basic and applied research, for purposes ranging from signal transduction and protein interaction studies, over viral packaging, to rapid smallscale protein expression for structural biology and recombinant pharmaceutical production. For complete details, download the datasheet via the get pdf button below. With the advent of viral gene therapy, 293 cells were used to produce the e1adeleted ad vectors, and they became one of the few cell lines approved by the fda for virus production for clinical trials.
Fetal cells from abortions are being used to flavor commercial foods and drinks or in the production of a mysterious substance known as adrenochrome. The history of hek293 and its use in vaccines and food testing. It is easy to grow and transfect and have been widely used for cell biology research and also used by the biotechnology industry to. Year of origin october 2008 references graham fl, et al. These may be the cell type of origin of 293 and other ad transformed cell lines. Recombinant protein expression for structural biology in hek. The 293ebna cell line is intended for use with vectors containing an ebv origin of replication oripyoung et al.
The hek293 human cell lineage is widely used in cell biology and biotechnology. Hek halo ce editing kit used to edit and create maps for the halo custom edition game only. Store frozen cell vials in the liquid nitrogen vapor phase. Invitrogen lipofectamine ltx reagent is a proprietary, animal origin free formulation for the transfection of dna into eukaryotic cells with low cytotoxicity. The scattered twelve tribes of hek293 biomedical and. Optimization of hek 293 cell growth a the effect of 10% fbs supplementation in commercially av ailable serumfree media, b growth kinetics of hek 293 cells in 3 different serumfree proteinfree formulations bi, bii and biii in the presence or absence of a predefined mixture of supplements, c effect of recombinant proteins or synthetic lipid mix on hek 293 cell growth, d. Genome dynamics of the human embryonic kidney 293 lineage. The first recombinant human coagulation factor viii of human origin. Hek halo ce editing kit used to edit and create maps for the halo custom edition game only file name. This situation may be solved by adopting more efficient production methods. Hek 293 cells are useful for many transfection experiments, particularly the propagation of adenoviralbased and retroviralbased vectors. A hek293derived cell line had been engineered harboring a single target locus with open reading frames encoding the livingcell reporter proteins lacz and egfp, the latter translationally.
Hek 293293human embryonic kidney cells 293,hek293hek293. The medium is complete and readytouse, with no supplementation required. The 293 ebna cell line is intended for use with vectors containing an ebv origin of replication oripyoung et al. Purchase 293 hek293 cell line for research and science from. Characteristics of a human cell line transformed by dna from human adenovirus type 5. It was therefore necessary to confirm the epithelial origin of the hek293 cells. Origins of the hek293 cell line hek293 is a cell line derived from human embryonic kidney cells grown in tissue culture. Download the raw data complete genomics the whole complete genomics package including sequencing reads and analysis from complete genomics is available on the european nucleotide archive ena repository, accession number prjeb3209. Looking for online definition of hek293 or what hek293 stands for. Hek 293 cells were generated by transformation of human embryonic kidney cell cultures hence hek with sheared adenovirus 5 dna, and were first described in 1977 graham et al.
Preferential transformation of human neuronal cells by human. Preferential transformation of human neuronal cells by. Material safety data sheet for cell cultures biosafety. The modal chromosome number was 64, occurring in 30% of cells. Hek 293 is used to deliver the lentivirus gene in dr yamanakas adult skin cell reprogramming, as a cell line and reagent for testing many research products and in the drug xigris. Hek halo editing kit for halo ce custom edition section. Evaluation of hek293 cell growth and adenovirus productivity. Hostrange mutants of adenovirus type 5 defective for growth in hela cells. Hek293 cells display a variety of features of the late distal tubule. An embryonic adrenal precursor cell therefore seems the most likely origin cell of the hek 293 line. Gpr124 is an orphan member of the adhesion gproteincoupled receptor family that is expressed in ecs and is essential for cns angiogenesis and barrier formation via an unknown. The 293 cell line was created by frank graham over 25 years ago by transfecting human embryonic kidney hence the name hek 293 cells with sheared ad5 dna.