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Why are HEK 293 cells used for transfection?

Why are HEK 293 cells used for transfection?

Advantages of HEK293 cells Firstly, they are very easy to grow and to maintain, with high reproducibility, which makes them preferable over other less-robust and slow-growing cell lines. Furthermore, they are very efficient at protein production and accessible for transfection.

How do you transfect cell protocols?

In a standard transfection protocol, the cells are plated on day 1, transfected on day 2 and assayed on day 3 or 4. In a reverse transfection protocol, cells are added directly to a plate containing the transfection reagent/DNA mix and assayed on day 2 or 3.

What is the difference between HEK293 and HEK293T?

The key difference between HEK293 and HEK293t is that HEK293 is an immortalized cell line derived from the embryonic human kidney that is transfected with sheared human adenovirus type 5DNA while HEK293t is a daughter cell line derived from HEK293 original cell line that is transfected with a plasmid vector carrying …

What does HEK293 stand for?

human embryonic kidney cells
What Is HEK293? HEK293 are immortalized human embryonic kidney cells. Originally isolated in the 1970s by Alex Van der Eb, a Dutch biologist, it was postdoc Frank Graham who transformed the cell line with sheared adenovirus 5 (Ad5). The ‘293’ refers to the fact that it was Graham’s 293rd experiment.

What have HEK293 cells been used for?

HEK 293 cells have been widely used in cell biology research for many years, because of their reliable growth and propensity for transfection. They are also used by the biotechnology industry to produce therapeutic proteins and viruses for gene therapy as well as safety testing for a vast array of chemicals.

How do you transfect HEK293 cells?

The protocol for a 24-well transfection reaction with HEK293 cells is here:

  1. Plate 10,000-15,000 HEK293 cells per well in 0.5 ml of complete growth medium 12-24 hours prior to transfection.
  2. Wash with 1xPBS and add 0.5 ml of fresh growth medium.

Do I need to change media before transfection?

It depends when you plate the cells for the transfection. If you plate the cells one day before transfection, then it may not be necessary to replace the medium. Off course, if you plate cells several days before the transfection, then it may be a good idea to change the medium before transfection.

What is cell line HEK293?

293 [HEK-293] is a cell line exhibiting epithelial morphology that was isolated from the kidney of a human embryo. This cell line can be used in industrial biotechnology and toxicology research. It has applications in efficacy testing and viruscide testing. Homo sapiens, human.

Can you transfect cells twice?

All Answers (7) Yes, it can be transfected, in principle.

What is HEK 293 transfection?

Transfection Information Human embryonic kidney 293 (abbreviated HEK 293 or HEK cells) are a cell line that was originally cultured from the kidney cells of a human embryo placed in tissue culture. These cells were transformed with sheared adenovirus DNA in a laboratory in Leiden, the Netherlands.

What is the subculture protocol for HEK293 cells?

Subculture Protocol for HEK293. An easier method to subculture cells is by centrifuging and resuspending in fresh medium: Transfer cell suspension to centrifuge tube Rap side of emptied flask to remove any semi-adherent cells Resuspend newly dislodged cells in 5 mL medium and aspirate with a serological pipette to disperse clumps Transfer…

How to reduce the cytotoxicity of HEK293 cells after transfection?

If the viability of HEK293 cells being transfected is affected at 16-24 hours post-transfection, changing the growth medium and eliminating redundant exposure of cells to transfectant can reduce the level of cytotoxicity Transfection. Cells and Molecular Biology Research Methods, Protocols, and Lab Techniques

How do you calculate viable cells from HEK 293 cells?

% viable cells = [ 1 – (number of blue cells/number of total cells) ] * 100 Production of P-glycoprotein: HEK 293 cells were transfected to transiently express P-glycoprotein with a histidine tag to study the phenomenon of drug efflux.