Hello,

I am not familiar with the specific cell type but it sounds like they are less viable. That can derive from a contamination (e.g. mycoplasma).

I would try to sterilize the incubator & hood take a new medium and defrost a new batch.

I hope that will solve the problem.

Good luck.

Thanks for your answer but I've already confirmed the cells are mycoplasma free, made new media and thoroughly cleaned out the hoods and incubator I use.

Hi there,

As Ariel mentions, it sounds like the cells are not completely healthy. How are they growing in normal conditions (ie not in chamber slides)?

I'm not familiar with this cell lines, but some cell lines benefit from an attachment coat prior to seeding. Perhaps you could look into this?

Thanks for your replies:

@Halla, the cells seem better in plastic well plates but I still experience a similar problem. I have been coating the chamber slides with Matrigel to aid in attachment but  the majority of cells still seem to have lifted off.

@Zoë the cells were at day 4 (out of 7 days) of the BDNF differentiation protocol and so were differentiating nicely with the majority of cells having developed long neurites  and connecting with eachother, it was only when I've changed the media that they seem to have detached. I will stain the cells with PI to see if they are still alive

The next method I will try is to differentiate the cells both in plastic and chamber slides at the same time and rather than taking media off of the cells, I will add a double-concentrated amount of BDNF in media equivalent to what's in the well in case it's just a shearing effect I am experiencing from the pipette when removing the media

Try washing slides, then poly lysine coating ...

Did you coat with PLL?