My lab is studying neuroprotective effect towward Sh-SY5Y cells. However, when I seed the Sh-Sy5Y cells in Sigma 96 well plate coated with PDL (cls3842), 3 days later, all Sh-SY5Y cells clumped together forming small lumps. In normal, they should be divided indivudally, equally spread inside the wells. This abnormal phenomenon does not appear when we used in a 96 well plate without PDL.
It surprises me since I read papers that using SH-SY5Y cells coated with PDL before.
I am wondering have you heard of any similar observation, and do you have any suggestion to prevent this?
Or do you guys have any suggestion for how to do pdl toward plates?
Thanks!
Thomas
Are you sure there was no other diference between your two experiments?
Well, on wiki— "SH-SY5Y cells can form mounds of undifferentiated cells, which then spread differentiated cells into the surrounding area. This growth formation can be referred to as the "over-easy formation""
And I found that if SH-SY5Y were seeded by less than 1:8, they'd like to clump together.
And I recommend seeding ratio 1:3 to 1: 5.
https://en.wikipedia.org/wiki/SH-SY5Y
Thanks for your help!. Will differentiated and undifferentiated cells react differently toward the PDL? I am using undifferentiated shsy5y.
Hi,
free (unattached) PDL can be relatively cytotoxic. Therefore, after coating you should thorougly rinse the plate with buffer/Adest. Apart from that 96 well plates are not ideal to yield an even distribution over the surface. A certain degree of aggregate formation is quite normal in undifferentiated SH cells.
Good luck!
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