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

Figure 8.

Figure 8

ALDHhiCD44+CD24− and ALDHhiCD44+CD133+ breast cancer cells demonstrate enhanced tumorigenicity and metastatic growth in vivo following mammary fat pad injection. Cells were isolated by FACS as described in Figs 3 and 4 and injected into the right thoracic mammary fat pad of female NOD/SCID‐IL2Rγ null mice using an established model of spontaneous metastasis (5 × 105 cells/mouse; 4 mice/cell population). At 7 weeks (MDA‐MB‐231) or 12 weeks (MDA‐MB‐468) after injection, mice were killed and assessed for metastatic burden in the lung and elsewhere. Tissue sections were subjected to haematoxylin and eosin staining (five random sections/tissue/mouse), and the incidence and extent of metastasis was determined in a blinded fashion. (A) Primary tumour growth kinetics following mammary fat pad injection 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). Data are presented as the mean ± S.E.M. *= significantly different tumour size than respective ALDHlowCD44low/− subsets at the same time‐point (P < 0.05). (B) Quantitative analysis of spontaneous lung metastasis tumour burden (mean% of lung occupied by tumour) following mammary fat pad injection 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). Data are presented as the mean ± S.E.M. δ= significantly different than respective ALDHlowCD44low/− subsets (P < 0.05). (C) Incidence of spontaneous metastatic growth in lung and extrapulmonary tissues following mammary fat pad injection 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).