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. 2008 Aug 4;13(8b):2236–2252. doi: 10.1111/j.1582-4934.2008.00455.x

Figure 6.

Figure 6

ALDHhiCD44+CD24− and ALDHhiCD44+CD133+ breast cancer cells demonstrate enhanced adhesion, migration and invasion in vitro. (A) Cell adhesion of ALDHhiCD44+CD24− versus ALDHlowCD44low/−CD24+ cells isolated from the MDA‐MB‐231 cell line (left panel); and ALDHhiCD44+CD133+ versus ALDHlowCD44low/− CD133− cells isolated from the MDA‐MB‐468 cell line (right panel). Cells were isolated by FACS as described in Figs 3 and 4 and plated onto sterile 96‐well non‐tissue culture plates treated with either 10 μg/ml of fibronectin (MDA‐MB‐231 cells), 5 μg/ml of vitronectin (MDA‐MB‐468 cells) (black bars) or PBS (negative control) (white bars), using 1 × 104 cells/well in triplicate wells for each sorted cell population. Cells were allowed to adhere for 5 hrs and adhered cells were quantified by manual counting of five high‐powered fields (HPF) per well. Data are presented as the mean ± S.E.M. *= significantly different than respective population of ALDHlowCD44low/− cells adhered to vitronectin or fibronectin (P < 0.001). (B) Cell migration and (C) Cell invasion of ALDHhiCD44+CD24− versus ALDHlowCD44low/−CD24+ cells isolated from the MDA‐MB‐231 cell line (left panels); and ALDHhiCD44+CD133+ versus ALDHlowCD44low/− CD133− cells isolated from the MDA‐MB‐468 cell line (right panels). Transwells (8 μm) were pre‐coated with either gelatin (migration assays; 6 μg/well) or Matrigel (invasion assays; 10 μg/well) and FACS‐isolated cells (3.5 × 104 cells/well; n= 3 for each cell population) were allowed to migrate or invade for 24 hrs towards chemoattractant media (5% foetal bovine serum) (black bars) or control media (0.01% bovine serum albumin) (white bars). Migrated or invaded cells were quantified by manual counting of five HPF per well. Data are presented as the mean ± S.E.M. δ= significantly different than respective population of ALDHlowCD44low/− cells migrating towards FBS (P < 0.001). φ= significantly different than respective population of ALDHlowCD44low/− cells invading through Matrigel towards FBS (P < 0.001).