Asked 17th Aug, 2012

What is the difference between Hek293 and Hek293T cell lines?

Who knows the difference between these 2 cell lines: Hek293 and 293T? I've used 293T for virus production.

Most recent answer

4th Jan, 2022
Slawomir Lukomski
West Virginia University
Does someone knows whether constructs made in pFUSE vectors are compatible with expression in HEK 293-series cells?

Popular Answers (1)

17th Aug, 2012
Michel Ouellet
Laval University
The 293T cells are derived from 293 cells but stably express the SV40 large T antigen which can bind to SV40 enhancers of expression vectors to increase protein production.
56 Recommendations

All Answers (26)

17th Aug, 2012
Michel Ouellet
Laval University
The 293T cells are derived from 293 cells but stably express the SV40 large T antigen which can bind to SV40 enhancers of expression vectors to increase protein production.
56 Recommendations
18th Aug, 2012
Luis Quintino
Lund University
293T are also neomycin resistant due to the presence of a neomycin resistance cassete together with the SV40 Large T .
9 Recommendations
20th Aug, 2012
Manoj Balakrishna Menon
Indian Institute of Technology Delhi
In addition to the comments above, the presence of T-antigen in 293T cells could help in episomal maintenance of SV40 origin containing vectors. In case of viral production and for other transfections 293T is now preferred since they are normally better transfectable.
6 Recommendations
20th Aug, 2012
Lijo Cherian
University of Copenhagen
Hek293T is an inbreed of Hek293 along with Tantigen of SV40, which gave them also added neomycin resistance and Temperature sensitive. Now this T-antigen can also bind to several proteins which inhibit cell cycle. So the cell is forced entering into more number of cell cycle. This adds on to increase the amplification of the transfected plasmid thus increasing the efficiency of transfection, higher vector production and higher transduction efficiency.
For your work purpose ie. virus production, you also need to check for particular clones of hek293T (usually available from ATCC) expressing special vectors which will lead to higher titre production.
Good luck!
12 Recommendations
20th Aug, 2012
Alasdair Russell
University of Cambridge
Hi Priscila
Just to add to Michel and Manoj's comments above, there are several variant 293T cell lines floating around in laboratory tissue culture hoods. While some may be resistant to neomycin and also carry ts mutants, others do not. Also, the LT's are driven off different promoters in some of these lines. It's also worth noting that there are subsequent derivatives of 293T's such as the 293FT cell line (which are a fast growing 293T isolate that (supposedly) increases lenti titre's substantially over that of the parental 293T line).
Good luck.
28 Recommendations
21st Aug, 2012
Jakob J Gutzmann
Eunice Kennedy Shriver National Institute of Child Health and Human Development
I would like to add to the above comments that in my experience the 293Ts are a little bit harder to handle. The detach quite easily from the culture flask and it is therefore especially important to split them regularly and not to let them overgrow.
Frohes Forschen and good luck with your experiments.
5 Recommendations
4th Sep, 2014
Appalaraju Jaggupilli
Syngene International Ltd.
I am trying to make stable cell lines with HEK293T cells using Geneticin for selection. From the above comments I understand that this cell line in particular is resistant to Neomycin/geneticin. Can anyone plz give me a good reference indicating the development of neomycin/geneticin resistance in HEK293T cells, so that I can move on to other cell line basing on a stronger evidence for my option.
14th Sep, 2014
Piyush Chudasama
Sat Kaival Hospital Pvt Ltd
Completely different question but can this cell line be used as normal cell line??
23rd Apr, 2015
Youhong Su
Nicole, The 293-EBNA cell line stably expresses the Epstein Barr Virus (EBV) EBNA-1 gene from pCMV/EBNA. Expression of the EBNA-1 gene is controlled by the CMV promoter and is high-level and constitutive. The 293-EBNA cell line is intended for use with vectors containing an EBV origin of replication (oriP)(Young et al., 1988). Vectors containing an oriP region can be maintained episomally in 293-EBNA cells. Please see the attached file from Lifetechnology for your reference.
3 Recommendations
25th Sep, 2015
Dennis Karthaus
Sartorius Xell GmbH
In our experience HEK cells containing the EBNA gen allow higher transient transfection efficiencies than 293T cells. Some literature also indicate that the EBNA gen enhances the transport of plasmids containing an oriP into the nucleus.
1 Recommendation
Deleted profile
Hi all.
I have one question regarding protein expression level of my transfection in 293t cell.
In my case the vector has two promoters, CMV and SV40, and I transfect this plasmid into 293t. I found the expression level is really huge.
However, I am trying to decrease overexpression in my experiment.  I would like to decrease the expression level.  Is it better for me to choose 293 instead of 293t?
Will SV40 make the expression much more enhanced in 293t?
3 Recommendations
17th Jun, 2016
Shravan Singh
Defence Research and Development Organisation
NK1R receptor is absent in the 293T ,whereas 293T cells are resistant to SP even at milimolar concentrations. 
1 Recommendation
14th Sep, 2017
Justyna Augustyniak
Mossakowski Medical Research Institute Polish Academy of Sciences
The 293T cell line is:
-          derivative of human embryonic kidney 293 cells,
-          a highly transfectable,
-          contains the SV40 T-antigen,
-          competent to replicate vectors carrying the SV40 region of replication.
-          It gives high titers when used to produce retroviruses.
3 Recommendations
25th Dec, 2017
Md Sahadat Hossain
Macquarie University
Basically HEK293 cells are the parental line for HEK293T. Here the letter "T" implies that it expresses the Large T antigen that is essential for replicating plasmids containing an SV40 origin of replication to high copy number in the transfected cell.
16 Recommendations
27th Mar, 2018
Joel P Joseph
Indian Institute of Science
@ Piyush Chudasama: HEK 293T is not a recommended normal. HEK293 can be used as a normal cell line. However, it is better to avoid them in case you are performing any apoptotic assays.
28th May, 2018
Antonio di stasi
University of Alabama at Birmingham
It looks like the 293T (ATCC® CRL-3216™) do have the neo Resistance, and I wonder why on the specification sheet there is no mention to periodically passage them in Neo.
-Also some protocol call to transfer them at 32C 24 hours before harvesting the sup b/c the retrovirus is more susceptible to 37C.
1 Recommendation
8th Aug, 2018
Dhiru Bansal
University of Cambridge
Joel P Joseph HI Joel , Why do you say to avoid HEk293 cells for performing apoptotic assays? Just curious because thats what I was planning for my experiment.
19th Aug, 2018
Joel P Joseph
Indian Institute of Science
@Dhiru Bansal I had read that they are resistant to apoptosis, so the read out may not be very good.
11th Jan, 2019
Abdulkareem Olarewaju Babamale
University of Ilorin
Does 293 differ from 293T cells. Because during my study, i got that 293 is fibroblast and 293T is epithelial cells? I need some clarification
24th Apr, 2019
Andreas Eisenreich
German Federal Institute for Risk Assessment / Bundesinstitut für Risikobewertung
HEK293T cells are optimized cells generated from HEK293 cells by stable transfection with a SV40 expression construct. They are standard cells often used as standard cells for transfection (overexpression, knockdown etc.) experiments.
5 Recommendations
Deleted profile
I have a follow-up question.
Is it worth to clone SV40 ORI into my expression vector used to transfect 293T cells??
As far as I understand having an SV40 ORI in your mammalian expression vector results in replication of your plasmid when the transfected 293T cells replicate.
Is there any data on how much this increases protein yield?
Are there limitations in the size of the plasmid?
Can or should you transfect the 293T cells at a lower confluency?
Any help would be greatly appreciated! Links to websites or papers are very welcome too!
1 Recommendation
15th Dec, 2020
Myriam Ben Khelil
University Bourgogne Franche-Comté
Hi Joel P Joseph thank you for this information about avoiding apoptotic assay with HEK293 cell line. Do you have any reference?
12th Oct, 2021
Fida Azar
French National Institute for Agriculture, Food, and Environment (INRAE)
Is it adviced to use the HEK293 or HEK293T cells for drug metabolism or drug screening experiments? if not is there any other "normal" human cell line that can be used?
Thank you

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