Hello everyone,

I'm working with D341 medulloblastoma cells and trying to assess their clonogenicity. These are suspension cells, and I’ve been using Poly-D-Lysine (PDL)-coated 6-well plates. While the cells initially attach, they often detach during media changes, which I typically do every 3–4 days based on cell density.

Interestingly, in a recent experiment where the initial seeding density (150K cells/well) was high and the culture reached 100% confluence, the cells attached well and remained stable. This makes me think that cell-cell interactions at high density might contribute to better colony integrity.

I’m also considering switching to a soft agar colony formation assay, as it may be more suitable for these cells. Has anyone used agar-based methods with D341 cells or similar suspension lines? If so, I’d appreciate any protocols, especially for soft agar or other methods that maintain colony structure over time.

Additionally, if you have a colorimetric assay recommendation for D341, I’d love to hear it.

Thank you,

Mohamad Rahmdel