Hello Ramneet Kaur
You may perform mild pipetting up and down, the cell clumps are easily dissociated, while mammospheres, to be efficiently dissociated, need to be enzymatically or mechanically disaggregated.
The cell density affects the ability of the cells to grow as mammospheres, making it difficult to distinguish cell aggregation from mammosphere formation. Due to the mobile nature of cells in suspension, cells drift and collide, leading to an aggregation tendency that is proportional to the cell density. This is problematic because clonality is an integral concept to the mammosphere formation. Mammospheres should arise from a single cell to effectively measure stem-like propagation. So, using low density plating will help reduce problems of cell aggregation.
I have attached two papers for your reference which may help you in your project. Please go through them.
https://www.cell.com/iscience/pdf/S2589-0042(18)30127-5.pdf
Article Determining Mammosphere-Forming Potential: Application of th...
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