I have been culturing human astrocytes derived from brain samples. Do not have any problem growing them on T25, T75, and multi-well plates (Poly D lysine coated). But, when I grow them on coverslips coated with PDL, astrocytes detach and I am barely left with any cells on the coverslips. This happened many times despite troubleshooting. Astrocytes adhere very nicely on coverslips coated with PDL, but they detach easily after PFA fixation and washes with PBS. All the steps have been performed very gently to avoid cell detachment. But nothing worked. So, I seeded human astrocytes directly on the chambered slides (without PDL coating), and I faced the same problem. I couldn't figure out where the problem is. I checked many papers that mentioned growing primary mouse and human astrocytes on PDL-coated coverslips.

I request experts who worked on human astrocytes to kindly suggest what I should do.

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