We analyse the mammosphere-forming ability of cells derived from clinical samples. Prior to their use in the mammosphere assay, epithelial cells (EP) first need to be extracted from such samples.
So, I need Know How to extract EP from Breast tumor samples ?
ISET (isolation by size of epithelial tumor cells) method is generally used technique.
Moreover, Molecular and immunomorphological characterization can also be done with this method.
you can also do FACS for EpCAM (epithelial cell adhesion molecule)-positive cells. so you collagenase-treat your tissue, stain it for EpCAM using a protocol for flow sorting, and use a FACS cell sorter to isolate those cells.
Even easier is to use EpCAM antibody-labelled magnetic beads and separate them magnetically.
After collagenase treatment better do CD45 stain which is a pan leukocyte marker and elute the non CD45 cells in that again you can stain for EpCAM. This way you will ignore false EpCAM positive cells.
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