Dear Dr. Mona Widhe

You should culture spheroids in less than 200µL of culture medium (100µL) in 96-well plate as it better mimics the in vivo conditions. If you have a smaller volume of medium, it will help encourage the cells to aggregate and form free-floating spheroids by reducing the likelihood of cells adhering to the plate surface. Spheroids are formed when cells are seeded in a low-adhesion environment. You should use ultra-low attachment plates as it will help prevent cell adhesion, allowing cells to form spheroids in suspension.

Also, smaller volume of medium, better reflect the nutrient and oxygen gradients that exist in tissues, which is important if you want to study cell behavior and drug response.

Regards,

Malcolm Nobre

Dear Dr. Malcolm Nobre,

Thank you for your reply and explanation. I will try this in coming experiments.

Best,

Mona