Cell passage

Discussion of all aspects of cellular structure, physiology and communication.

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Endothelial-cell
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Cell passage

Post by Endothelial-cell » Wed May 16, 2007 7:33 am

Hi all...

May i know what is the ideal passage for endothelial cell, to be use on cell culture? And why is it so? The ideal cell passage varies between different cell type? Thanks!

Cheers

blcr11
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Post by blcr11 » Mon May 21, 2007 4:11 pm

It's been a long time since I've done any tissue culture, so I may not be the best person to answer this kind of question, but there may not be a single optimum. The typical thing is to plate them at some density--1E3 or 1E4/ml or per plate (I don't remember which anymore)--and let them grow back to confluency. This may take anywhere from 3-7 days--possibly longer. At confluency, you usually treat the dish briefly with trypsin to release the adherent cells, collect the cells, dilute in fresh media and replate at whatever density you're using. If you're using "fresh" non-imortalized cells, you'll probably need to use supplemented media of some sort--but I think there are better options available now than when I did cell culture. I would recommend trying to find a local friendly tissue culturist for advice on those kinds of issues.

konstantin
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Post by konstantin » Fri Jun 01, 2007 3:54 am

hello, I am working with HUVEC (human umbolical endothelial cell)
actually this cell culture is very sensitive to any stress that's why you have to care them very well....
previous comment is ok, in order to do subculture it is neccessary to use tripsyn, but in my case there is one very important moment....they will not adhere to the buttom of petri dish by itself.....you have to coat buttom with 1% of gelatin....it make them to easy adhere...
and our lab..use HYQ 199/EBSS medium from HyClone....as well as content of medium is a little different for example instead of 10% I use 20% of FBS, heparin, ECGS, and PS.....
so I hope this information will be useful for you
if you have additional questions don't hesitate

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Post by Endothelial-cell » Thu Jun 07, 2007 1:20 am

That great! Thank!....as right now im using a substrate...to study how well EC adhere to the substrate...I would like to find out more about ur work....ur using 1% of gelatin, and do u mean u add in FBS, heparin, ECGS, and PS? Instead of buying a complete medium? Thanks for ur help!

konstantin
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Post by konstantin » Thu Jun 07, 2007 5:01 am

yes yes you are right, we add FBS PS and heparin as well as ECGS instead to buy complete media....
good luck in anyways....I am glad to help you...

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Post by Endothelial-cell » Thu Jun 07, 2007 5:08 am

Do u mind if u can give me the product code from HyClone? Then i can check it out. thanks!

konstantin
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Post by konstantin » Wed Jun 13, 2007 7:07 am

the catalog number of HyClone media is SH30253.01

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