I am experiencing difficulties seeding SH-SY5Y cells on Poly-D-Lysine or Poly-L-Lysine treated coverslips.After 2-3 days, the cells either detach or fail to attain their typical morphology. The attached cells exhibit clumping or clustering. Is there any solution for this issue?
Samir Ranjan Panda
Hi Deepak Mukundan K V
Please check the following protocol and suggestions.
- Sterilization: Sterilize coverslips by autoclaving or using another appropriate method.
- Coating: Prepare a solution of PDL or PLL (typically 0.1 - 1 mg/mL). Incubate coverslips in the solution for 15-30 minutes at room temperature. Gently rinse coverslips with sterile water or PBS to remove excess PDL/PLL.
- Drying: Air-dry the coverslips in a sterile hood for at least 30 minutes to ensure complete drying. Some protocols recommend additional drying steps like overnight incubation.
* To prevent the cells from forming clumps I would advise you to mix the cells thoroughly during resuspending so as to obtain individual cells.
* What is the % of FBS in your plating medium? You can increase the conc of FBS upto 15 % to enhance the cell attachment.
* Are the cells detaching post differentiation?
Thanks,
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