I am trying to do mammosphere formation assay. I want to ask if we can use non adherent (suspension culture) plates in place of Ultralow attachment plates. ULPs are expensive and I am looking for an alternative to do my experiment.
I think you can use non-adherent plates with minimal or lower poly o lysine coating for achieving low adherency in the plates. You can try optimizing the coating concentrations and check if your requirements are met or not.
Hii Shivani
Thanks for your suggestion.
It appears that non adherent plates still support some level of cell attachment through non specific interactions (hydrophobic or ionic). I tried to coat the non adherent plates with 2% agar to prevent any sort of hydrophobic or ionic interactions between the cells and the surface of the plate. This kept the cells in suspension while allowing their consistent growth to form spheres when cultured in serum free DMEM/F12 media supplemented with growth factors (VEGF, FGF and insulin).
Oh, I thought you required Ultralow adherence plates instead of non-adherent plates for your experiments as used in 3D cultures. That is why I suggested o-lysin coating. Anyways seems like you are able to grow clumps in suspension culture as well.
Yeah. I cultured these mammospheres for 21 days. I could see the spheres growing in clumps in suspension cultures. The technique worked well. You can also check the following articles if you are interested:
Article Development of a novel and economical agar-based non-adheren...
Article Characterization of the stemness potency of mammospheres iso...