Hi Polina,

• After thaw, plate at higher density and keep ROCK inhibitor for 24–48 h; do only gentle half-media changes in the first 48 h.
• Reduce DNA-mediated clumping with a brief DNase I step post-thaw and use very gentle trituration; avoid over-handling early.
• Ensure coverslips are TC-treated and coatings are fresh/rinsed; add a laminin overlay to improve spread and neurite health.
• For more even adhesion and less clustering, use DendroTEK dPGA | Protease-Resistant Culture Matrix as the base layer (instead of PDL/PLO), then overlay with laminin.

Best of luck, Jason Hamlin

Source: I work at DendroTEK Biosciences. www.dendrotek.ca