How do you freeze HEK293 cells?

As cells are dispensed into cryovials, frequently and gently mix the cell suspension to maintain a homogenous solution. [Add 1mL in each vial using 10mL pipette] 9) Freeze the cells in the -80°C freezer in a controlled rate freezing container with isopropanol by decreasing the temp approximately 1°C per min.

How long does HEK293 cells to adhere?

Most HEK-293T cells double every 12–20 hours depending on conditions. Our cell doubling time is typically closer to 12 hours.

Why are HEK293 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 split a 293 cell?

Splitting Cells: Aspirate the medium from the flask. Wash the cells carefully with PBS to remove residual medium. Add 1-2ml of Trypsin Solution (equilibrated at RT) to the flask (150 cm2) and incubate at 37°C until cells have detached (1-2 minutes). Prepare a new flask with fresh medium.

What is the purpose of freezing cells?

Recovery Cell Culture Freezing Medium is a ready-to-use complete cryopreservation medium for mammalian cell cultures, containing an optimized ratio of fetal bovine serum to bovine serum for improved cell viability and cell recovery after thawing.

Where does HEK293 come from?

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.

How are HEK293 cells harvested?

HEK 293 cells were generated in 1973 by transfection of cultures of normal human embryonic kidney cells with sheared adenovirus 5 DNA in Alex van der Eb’s laboratory in Leiden, the Netherlands. The cells were obtained from a single, aborted or miscarried fetus, the precise origin of which is unclear.

What else is HEK293 used for?

HEK293 are frequently used for heterologous expression of cell membrane receptors and ion channels, and many studies have utilized this cell line to investigate the signalling pathways of G protein-coupled receptors (GPCRs) which are implicated in disease.

How many times can you passage HEK293 cells?

However, the number of passages – not only for HEK293T cells – frequently depends on specifics of the laboratory and/or the person propagting the cells. We propagate HEK293T cells twice a week at split ratio of 1:3 and use bdbiosciences cell culture material. Cultures should be confluent after 3 days.