Hello everyone,
lately I'm working with spheroids of primary mice intestinal stem cells. I'm trying to set up some 2D culture as well, trypsinizing the spheroids and seeding the individualized cells. I coated the 96 well plate from corning with 2% matrigel of the same brand; my cells are able to attach to the surface, but they die soon after. I've not a lot of experience with this type of cells, so I would ask if you have some suggestion to make them survive or if someone did it already before.
Thank you to everyone!
Hi Federica Quacquarelli,
I don't have a lot of experience with intestinal stem cells, however I do know that media formulation is absolutely critical, and it would be useful to know what you are using.
Best of luck!
Sam
Dear Sam Siljee ,
thank you for your answer. The medium we use is generally DMEM supplemented with PSG, EGF, Nogging and R-spondin. No serum
Hi Federica Quacquarelli
Just before plating the cells, gentle rinsing of the coated surface with medium may help. Higher concentration of adhesion substrates can be toxic to cells. You can try using medium without any growth factors and supplements while plating and then replace it with the regular medium once the cells have adhered, which usually takes 3-4 hours. Good luck.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining