18 replies to this topic

[polarka]

Hello,
do you have experience with TEER measurements on HaCaT cells? I measured it, but the values were the same for blanks as well (transwell without HaCaT cells had the same TEER value as Transwell with HaCaT cells). Do you know what could be the reason?
I was thinking that HaCaT cells were not grown as monolayer and therefore TEER could not be measured - because I think that TEER is possible to measure only on monolayer. Do you have explanation?
K

[chowyc2]

cotchy, on 23 April 2009 - 01:00 AM, said:

Hi all, a colleauge of mine has recently started to work with HACAT cells.

He bought them in a while ago from CLS (cell line company) at a low passage number and passaged them a few times and then froze them down in the - 80C (70 % DMEM, 20 % serum, 10 % DMSO) and then after a few days places them in liquid nitrogen

When he brings them up from the liquid nitrogen (in the proper recommended media - DMEM w/ 10 % serum ) for use the viabilty of the cells is always very low, around 2 - 5 % and thus it takes 2 to 3 weeks to get a T 25 confluent.

Does anybody have any suggestions as to what is causing the low viability of the cells or has anybody had the same problems and managed to overcome them?

A US company sells the same cells, here is the link [link]http://addexbio.com/...tdetail?pid=117[/link]. Check that out to see if they also come up with culture condition for this cell line.

[Chakchel]

For development of TEER, you have to incubate the HaCaT cells with medium that contains more calcium than the usual HaCaT medium to induce further differentiation. After a few days the tight junction network should be well-established.

[honeybeeresearcher]

Just started to grow the HaCaT cells since we bought it a month ago from addexbio [link]http://www.addexbio....tdetail?pid=117[link] and the cells grows wonderfully. Highly recommend their cells. Customer service is great too. Planning to buy another cell line from them.