Modulations in cell proliferation levels of MSDACs under alternated culture conditions. A) Representative microphotographs of random fields obtained from Operetta high-content confocal imaging showing BrdU incorporation levels in three different clones of MSDACs maintained in serum-free stem cell medium for three generations: SF-SCM-1(3G), SF-SCM-1(3G) cells transferred and maintained in growth medium with FBS for three generations (SF-SCM-1(3G) → GM-FBS(3G), or the MSDACs reintroduced in SF-SCM for three generations (SF-SCM-1(3G) → GM-FBS(3G) → SF-SCM-2(3G). Parental SH-SY5Y cells maintained in growth medium are used as controls. B) Histograms obtained from Columbus image analysis showing significant decline in BrdU positive cells in SF-SCM-1(3G) → GM-FBS(3G) MSDACs and regained and amplified cell proliferation in MSDACs reintroduced in SF-SCM (SF-SCM-1(3G) → GM-FBS(3G) → SF-SCM-2(3G). Brd-U, bromodeoxyuridine; MSDACs, metastatic site-derived aggressive cells.