Here is a summary of the info I gleaned from asking for recommendations for the best human cell line. These were my criteria:
For context, we currently use a number of human cell lines in the lab: HeLa, RPE1, HCT116, SKOV3; as well as many others in the past: HEK293, DLD-1, U2OS.
I consider HeLa to be the almost perfect cell line. They are nice for imaging (great for biochemistry too), easy to culture and transfect… unfortunately they have a weird karyotype, so gene editing is painful. Plus, there’s always pushback from people who say HeLa are strange and not very representative of a “real cell”… a discussion for another day perhaps. So what cell line could be better?
hTERT-RPE1
Retinal Pigment Epithelial cell line (hTERT immortalized)
https://www.cellosaurus.org/CVCL_4388
Nishit Srivastava @Nishit24
maybe RPE1
Dr Rafal Czapiewski @RafalEstrella
RPE1 are my favourite now!
Fernando M. Valbuena @FernandoMValbu3
I use RPE-1 cells regularly. They’re fantastic for imaging, aren’t tumor derived, and have a stable karyotype. I use Neon electroporation for transfection with very high efficiency even when using mini prepped DNA. Electroporation is a better method than lipid-based reagents.
Ewan MacDonald @EwanMac10292433
Have you tried RPE1 cells?
Andrii Gorelik @AndriiGorelik
With PolyFect you can get >50% transfection efficiency in RPE1
Miguel Rz @MickeRzO
RPE-hTERT 🥇
Dr CJ Houldcroft @DrCJ_Houldcroft
What about TERT’d fibroblast lines?
Dr. Ceri Fielding @CeriFielding
Not that easy to transfect by conventional means, unless you’re transfecting siRNA.
Stephen Terry @Stephen17822072
Hi Steve I think rpe-1 htert cells I got are human near diploid flat so great for imaging transfect well for dna /siRNA the ones I have are puro resistant only drawback
Francesca Bottanelli @Fran_Bottanelli
We successfully made KIs in MCF10A and RPE1 cells. They transfect well with electroporation (NEPA gene).
ARPE-19
Retinal Pigment Epithelial cell line (spontaneous immortalized)
https://www.cellosaurus.org/CVCL_0145
Costin Antonescu @CostinAntonescu
We use ARPE19 for many applications (different from RPE1). Great for membrane traffic imaging especially TIRF. Reasonable for transfection, but drawback is they don’t edit easily.
Daniel Chauss @dchauss
ARPE19 also has reported karyotype heterogeneity
https://pubmed.ncbi.nlm.nih.gov/29476476/
HFF
Human foreskin fibroblast
https://www.cellosaurus.org/CVCL_XB54
Tim Feinstein @TimNFeinstein
HFF cells have a lot of good properties and are more ‘stem-like’ than most cultured lines, but if you don’t like the transfection properties of RPE-1 cells then they might not work.
Caroline Clucas @Caroline_Clucas
HFFs?
WI-26/WI-38
Fetal lung fibroblast
https://www.cellosaurus.org/CVCL_V826
https://www.cellosaurus.org/CVCL_0579
Ishier Raote @ishier_raote
WI-26 or WI38?
Julian Nüchel (PhD) @JNuchel
That’s what I would suggest! My favorite cell line! Perfect for endocytosis, secretion and other intracellular trafficking pathways! Generated plenty of KOs and OE cells. Transfection worked best for me with Xtremegene HP from Roche/Merck
U2OS & COS-7
Bone sarcoma
https://www.cellosaurus.org/CVCL_0042
African green monkey kidney
https://www.cellosaurus.org/CVCL_0224
Hoang Anh Le (aka Anh), PhD @AnhHLe2702
U2OS or COS7 (monkey but human siRNAs and CRISPRs work for these cells)
Andy Moore @aaandmoore
I’ve had mixed success with human siRNA/CRISPR in COS7. I guess it’s dependent on the specific gene. I’ve switched to just designing things specifically for vervet, but then it’s kind of annoying to have a tool that only works for Vero/CV1/COS.
Hoang Anh Le (aka Anh), PhD @AnhHLe2702
I had one instance that the target siRNA didnt work for one gene in COS7, but luckily for other genes they worked. So yes, it is gene dependent.
Hoang Anh Le (aka Anh), PhD @AnhHLe2702
I see. I did have a bit of problem editing U2OS, but easier for COS7. I also heard SKOV3 cells are really easy to culture too. My labmate used SKOV3 and they seem to love them.
Jonny Nixon-Abell @AbellJonny
They aren’t I’m afraid…which is a shame because damn they look good! U2OS often aren’t diploid either but more stable karyotype. Could use the KOLF2.1 iPSC line, dont image so well, but meet your other criteria.
John Petrini @JohnPetrini
I would strongly advise against U2OS. They are extremely unstable karyotypically. See: PMC3400581
https://europepmc.org/article/PMC/PMC3400581
Tyson V. Sharp @tyson_sharp
U2OS? Massive nuclei and massive cytoplasm!
Jullien Denis @JullienDenis1
Try U2OS
Stem cells
David Drubin @DavidGDrubin
For the reasons outlined here (https://www.molbiolcell.org/doi/full/10.1091/mbc.e17-03-0183) consider WTC human stems. Great protocols and a large catalog of engineered cell lines available thanks to @AllenCellInst. You can differentiate these cells into many other cell types (e.g., fibroblasts, cardiomyocytes)
Dr Agathe Chaigne @AgChaigne
Human, not mammals? mESC are pretty stable karyotypically (am I making up a word?) and I see a lot less division defects than hESC. They are not that hard to culture/transfect/image.
Max Gutierrez @maxgabgut
Cell line and karyotype stable are not compatible I am afraid. As suggested by others iPSC are an option that will not solve all your problems. If you use them at the iPSC stage results can be different from a fully differentiated cell. Good luck!
HT-1080
Fibrosarcoma
https://www.cellosaurus.org/CVCL_0317
Graham Dellaire @DocDellaire
HT1080 are great, big flat cells for microscopy. They are not diploid but have a stable phenotype from the point of view of rearrangements but do spontaneously become tetraploid if abused.
Francesco Pasqualini @fspasqualini
We love them but they also do like to crowl on top of each other which is not ideal for all microscopy needs
Aoqun James Song @AJamesSong
How about HT1080?
MDA-MB-231
Triple-negative breast cancer TNBC
https://www.cellosaurus.org/CVCL_0317
Aurélie Dobric @AurelieDobric
I may be too late for answer but I haven’t seen one cell line in the answer below : MDA-MB231. They are gorgeous for live imaging.
Shem @Shem_Darius
Not sure it checks all those boxes, but look at mda-mb-231 cells if you’re into endosomes. They’re huge!
Sherif Ibrahim. MBBCH, MSc, PhD @444sherif
Some authors reported difficulty in their transfection, but they worked well in my hand. I am not sure about their karyotype but they are easy to culture, and I believe they are great for microscopy.
DLD-1
Colorectal cell line
https://www.cellosaurus.org/CVCL_0248
Janardan Gavade @janardangavade
Maybe Dld-1 cell line can be the answer!
Melinda Beccari @MelsNeurons
Many here in lab use DLD-1s! If I’m not mistaken they fit all the items on your list. Let me know if you guys have questions about them or want a vial. We’ve used SH-SY5Y and U2Os also but always with lentivirus or nucleofection, not transfection.
MCF-10A
Breast cancer epithelial
https://www.cellosaurus.org/CVCL_0598
Valentine Comaills @ValComaills
MCF10A, great to work with. Media a bit weird but genomically stable.
Francesca Bottanelli @Fran_Bottanelli
We successfully made KIs in MCF10A and RPE1 cells. They transfect well with electroporation (NEPA gene).
THERY Manuel @ManuelTHERY
and hard to transfect
Valentine Comaills @ValComaills
Lentivirus, retrovirus and siRNA were fine for me.. but nothing too agressive as they are ‘normal’ cells and don’t like to be messed up, and end up to sense it and stop to grow.
HCT 116
Colorectal cancer
https://www.cellosaurus.org/CVCL_0291
대학원생 달토끼 @BunnyTheHuman_
How about HCT116?
ChrisStaples @StaplesLab
As well as the mutator phenotype, they are not so beautiful in fluorescence microscopy.
대학원생 달토끼 @BunnyTheHuman_
Although its small size, we use hct116 for PALM imaging.
Graham Dellaire @DocDellaire
I would stay a million miles away from HCT-116 due to MLH1 mutations and hyper mutation phenotype. They are a different cell line every few passages. Nearly every gene can accumulate point mutations.
Graham Dellaire @DocDellaire
For future reference (see Davis et al., 1998 Cancer Res.) since both Cellosarus and Wikipedia fail to inform users that HCT116 are MLH1-deficient…
https://pubmed.ncbi.nlm.nih.gov/9485033/
MRC-5
Lung fibroblast
https://www.cellosaurus.org/CVCL_0440
Fausto Capelluto @FaustoCapelluto
Have you tried MRC5? In my experience, they are very good for imaging and easy to culture.
Samantha Lewis @SamLewisPhD
MRC5 have beautiful mitochondria but ER looks like cotton candy… IME
Only the lonely: lines with just one recommendation
A-549
Lung cancer
https://www.cellosaurus.org/CVCL_0023
SA @SSA07844173
How about A549 cells?
CAL-51
TNBC cell line
https://www.cellosaurus.org/CVCL_1110
Jez Carlton @JezCarlton
I use CAL-51, near diploid, transfect well, edit well, great for imaging, divide every day. TNBC line with I think activating PI3K mutation.
HIEC-6
Intestinal epithelial
https://www.cellosaurus.org/CVCL_6C21
Dave Boucher @dave_boucher
Hiec-6 cells . Intestinal epithelial cells, grow well , easy media (optimum 4%fcs + egf)
HaCaT
Keratinocyte
https://www.cellosaurus.org/CVCL_0038
Polina Lishko @PolinaLishko
HaCaT (human keratinocytes) are great for 1-3, but they are aneuploid. Not sure how stable their karyotype is.
IMR90
Lung fibroblast
https://www.cellosaurus.org/CVCL_0347
Samantha Lewis @SamLewisPhD
IMR90. Normal karyotype, easy to transiently transfect, and high editing efficiency. They do senesce after 15-20 passages though.
SUM159
TNBC cell line
https://www.cellosaurus.org/CVCL_5423
Jeeyun Chung @JeeyunC
We use SUM159 breast cancer cell line. Diploid, easy to KI and KO, okay to transfect, beautiful ER structure for imaging!
Ewan MacDonald @EwanMac10292433
We use SUM159. They have nice trafficking, near diploid.
HEK293
Kidney
https://www.cellosaurus.org/CVCL_0045
Tim Triche, Jr. @timtriche
Presumably HEK293T is no good for microscopy of the sort you’re doing? Otherwise it’s nice
Jurkat
T-cell
https://www.cellosaurus.org/CVCL_0065
Candice @candice_ab
Jurkat cells?
Summary
Inevitably the answers were a classic case of YMMV. One person’s idea of a cell line that’s nice for imaging can be quite different from someone else’s. On top of all this, a wonderful cell line which doesn’t express the protein you are interested in (or do the thing you work on) is no use to you at all.
Does the perfect cell line exist? The strongest contenders all had drawbacks.
- RPE1 were the most popular suggestion. They’re great but hard to transfect.
- U2OS great but karyotypically unstable.
- HCT116 again the genomic concerns raised by Graham are off-putting.
There were several lines that we haven’t used and I’m keen for us to try. CAL-51, HT1080 and stem cells are all on our list.
Thanks to everyone who replied, I learned a lot.
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The post title comes from “Line Up” by Elastica which opens on their eponymous debut LP.