1. Volume 15, Issue 12, Pages 2088-2093 (December 2007)
Oncolytic Adenoviruses Kill Breast Cancer Initiating CD44+CD24–/Low Cells 
Minna Eriksson, Kilian Guse, Gerd Bauerschmitz, Pekka Virkkunen, Maija Tarkkanen, Minna Tanner, Tanja Hakkarainen, Anna Kanerva, Renee A Desmond, Sari Pesonen, Akseli Hemminki 
Molecular Therapy 
Volume 15, Issue 12, Pages (December 2007)
DOI: /sj.mt
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Isolation of putative breast cancer stem cells. CD44+CD24−/low cells were collected from pleural effusions of breast cancer patients using magnetic beads. The expression of CD44 and CD24 in (a) unsorted, (b) CD44−CD24−/low, and (c) CD44+D24−/low cells was analyzed by flow cytometry.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
Stem cell features of pleural effusion cells. (a) Unsorted and (b) CD44+CD24−/low cells were stained with Hoechst and imaged by fluorescence microscope. A subpopulation of CD44+D24−/low cells exclude Hoechst dye (arrows). (c) The expression of stem cell markers oct4 and sox2 in CD44+CD24−/low–sorted cells was confirmed by semi-quantitative real-time polymerase chain reaction. Fat tissue–derived mesenchymal stem cells were used as a positive control (+) and β-actin as a control for total RNA. The pleural effusion samples are indicated with sample number. (d) Immunofluorescence staining of Oct-4 (red) and CD44 (green) expression in the CD44+CD24−/low cell population.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
Killing of CD44+CD24−/low cells with oncolytic adenoviruses. (a–d) Cell killing was measured with MTS assay (Promega, Madison, WI). Positive and negative controls were wild-type adenovirus Ad300wt and replication deficient Ad5luc1. VP = viral particles. The value for non-infected cells was set as 100%. Data are expressed as means ± SEM. (e) The absence of living cells following infection with oncolytic viruses Ad5/3-Δ24 and Ad5.pK7-Δ24 was confirmed by trypan blue exclusion 8 days after infection.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
Anti-tumor efficacy of Ad5/3-Δ24 against CD44+CD24low/−–derived tumors. (a) Injection of CD44+CD24low/− cells fresh from patients resulted in tumors in all animals (N = 10 tumors in five mice), while pre-infection (filled symbols, N = 4 tumors in two mice), prevented growth in all cases. Size of tumors detected only after skin removal was set as 50 mm3. (b) CD44+CD24low/− JIMT-1 cells formed tumors in all mammary fat pads (N = 6 tumors in three mice) while pre-infection (filled symbols, N = 4 tumors in two mice) prevented tumor growth. Treatment of established tumors with 3 × 109 viral particles of Ad5/3Δ24 (arrows) stopped tumor growth and prevented death of mice.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

6. Figure 5
Anti-tumor efficacy of oncolytic viruses against CD44+CD24low/−–derived tumors. Injection of 2 × 106 cells into mammary fat pads of non-obese diabetic/severe combined immunodeficient mice resulted in tumors in 14 days (N = 8 tumors in four mice/group, i.e., total 40 tumors in 20 mice). Viruses or just medium were then injected thrice weekly (arrows) and tumor size was calculated: length × width2 × 0.5. All viruses gave an anti-tumor effect when compared to medium only (all P < ).
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions