Hi all, can anyone recommend a mouse endothelial cell line that can be used in tube formation assays, that is easy to culture and can be passaged several times?
Hi Derek,
you can use H5V. They are not as good as HUVEC (which are anyway human) but they do the job.
Ornella
You can try with SK/HEP which is adenocarcinoma cell line i think and which has endothelial origin.But before you use please check whether they express endothelial specific marker vWF and even presence of weibel palade bodies.
One of my past students had tried bEND.3 cells from ATCC, its a mouse endothelial cell line, for matrigel assay. According to a series of experiments that were carried, tube formation was transient in the sense that formation tended to disappear after 12-18 hours. I am not sure if I would suggest this for routine experiments. I can provide more technical details if you are seriously interested.
I didn't have much luck with mouse primary endos either, however tissue angiogenesis in matrigel provided fairly reproducible results. If you are interested in the later and can afford mice, I can provide some technical details.
Thanks everybody for all the answers!
Ornella, thanks for the advice, they look pretty good, do you know who supplies them I cant seem to find a supplier online?
Thanks
Hi derek,
I actually don't know. I had them in my previous lab, but I can send an email to check where they got it. They work pretty well and they express all the Endothelial marker (I tried already) they are much better in early passages. (like Huvec).. just remember, to do the tube assay, you need to starve them for 1 at least hour before using them.
I'll let you know as soon as I get a reply.
Ornella
Hi Derek,
I searched and they were given to me by a collaborator, but I think you can easily have them. if you give me an email I'll write to you.
Ornella
Sorry I cannot recommend you a mouse cell line for that assay. In fact I found very interesting this post because we tried the assay with primary lung endothelial cells and it didn't work.
A previous comment by Ornella mentions: " … you need to starve them for 1 at least hour before using them". I'm curious: Why is that? According to my experience with HUVECs that pre-condicioning is not required.
All the best,
Juan Carlos
Huvec do form tubes very easily. H5V continue to proliferate if you don't give them a stimuli to differentiate. This why you starve them.
Ornella
Thanks for sharing yuor experiences. I would agree with Ornella in that regard, I would suggest even longer time for starvation without risking cells for detachment. I am sure Ornella has a good explanation for this, here is my take on the issue.
Whether you do or don't serum-starve cells, they will still produce some sprouts n tubes however the cells may not respond homogeneously, which would be a serious issue if you were to evaluate the effects of some drugs or factors relatively quantitatively. If you do starve them, then you are removing or at least minimizing the influence of several factors in the serum and paracrine factors from the cells themselves on the angiogenic potenital of the cells. In fact, we do take this pecaution for all cell-based 'functional' enzyme assays in general.
I have tried human lung microvascular endos from Lonza, they did respond to SDF and VEGF that was fairly reproducibe. I've never tried primary lung cells though.
This company provides many primary mouse endothelial cells.
Good lucks
http://www.cellbiologics.net/
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