Johan, To me it looks like your cells are simply dying. Did you perform a viability stain? When do you see this? Primary intestinal cells may require more to grow than only DMEM.

Hi mr Markus,

Actually, they are not dying but instead after 8 days of incubation in DMEM at 27 dgrees, they change morphology and spreading like in the picture. Should I characterize them?

This Looks like fibroblasts to me. They often overgrow the epithelia since they are less demanding regarding culture conditions.

• Oh fibroblasts?...should i continue culture them or discard?they are from the guts of tenebrio molitor