Morphometrics of the parental SH-SY5Y and metastatic site derived aggressive cells and the schematic representation of the experimental workflow. (A) Representative light microscope photographs of parental SH-SY5Y cells and metastatic site-derived aggressive cells (MSDACs) maintained in serum-free stem cell medium. Parental cells exhibited monolayer spreading with organized neurites (100 ×). MSDACs are more spherical, smaller in size (100 ×), and formed organized tumorospheres (10 ×). (B) Schematic representation of experimental workflow: Three well characterized individual clones of MSDACs were alternated between stabilized (three generations) microenvironment simulations with serum-free stem cell medium to growth medium with serum and reintroduced back to serum-free stem cell medium. Parental cells grown in routine medium and serum-free stem cell medium were included as controls. Cancer stem cell physiognomy, tumorosphere formation capacity, and transcriptional and translational rearrangements associated with phenotype conversions were assessed at the end of each phase and in controls.