Hello,
Does anyone have any experience culturing HEK-Blue hNOD1 cells from InvivoGen? I cultured them for 2 weeks now. I notice that they are way too easy to detach. I just moved the flask from the incubator to the microscope, but most of them already detached from the bottom of the flask. I was wondering is that normal for this kind of cells? If not, do you have any suggestion how to fix this issue, please? Thank you very much
Panjaporn
Hi Chaichana,
I have worked with HEK-blue hTLR9 (SEAP reporter) also from Invivogen. But I guess they are in same system.
I have some experiments that I would like to share
HEK is always maintained at 50-60% confluence before seeding. If too confluent, you tap the flask, and it detaches
You better freeze them with large amount of vial and discard it in each single use. The result would be stable. Please do not keep culturing cells for the repeat and next use.
Regarding your hNOD1, you better optimize the seeding number for your assay, especially flourescent assay. As you get the right seeding number, you would get the significant result. But keep in mind that, we should use the black plate to minimize the reflection between wells and the wall of plate. Those can contribute to the background noise
As you add the substrate, try to measure kinetically and both from bottom and top of plate. Try to get the read earlier than the protocol recommending, and analyzing right away so that you can estimate the time to get the read on the next times.
Hope those helps
Good luck
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