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Blood, Vol. 113, Issue 22, 5423-5433, May 28, 2009
Toward a stem cell gene therapy for breast cancer
Blood Li et al.
113: 5423
Supplemental materials for: Li et al
Files in this Data Supplement:
- Figure S8. Distribution of mouse HSC-progeny after transplantation into myeloablated neu-tg mice (JPG, 528 KB)
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(A) Mouse Y-chromosome FISH on female (control) bone marrow cells and female recipients that received male HSCs (6 weeks after transplantation). Y-chromosomes appear as pink dots. (B) Mouse Y-chromosome positive cells in organs of transplant recipients. In intestine, Y-chromosome–signals co-localized with CD45 signals. (C) Distribution of RV-bGal–transduced HSCs in recipients’ bone marrow, lung, and intestine. bGal expression was visualized by X-Gal staining and appears in blue.
- Figure S9. Larger magnification of images in Fig. 5 (JPG, 515 KB)
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The size bar is 100mm. (A) Pink dots: mouse Y-chromosome, blue: nuclei; (B) green: mouse CD45, blue: laminin; (C) blue: bGal expression, counterstaining: hematoxyin; (D) green: CD45, red bGal.
- Figure S10. Blood cell analysis in recipients of mouse HSCs transduced ex vivo with LV-Rlx (JPG, 318 KB)
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Mouse HSCs were transduced ex vivo with LV-Rlx and transplanted into irradiated neu-tg mice. Six weeks later, MMC cells were injected subcutaneously. Three days after MMC transplantation mice received Dox. Control mice did not receive Dox. Three weeks after MMC cell transplantation, blood was collected an analyzed. N=5. wbc-whole blood cell count; NE-neutrophils; LY-lymphocytes; MO-monocytes; EO-eosinophils; RBC-red blood cell count; HB-hemaglobin; HCT-hematocrit; PLT-platelet; BA-basophils.
- Figure S11. CD3+ cells in mouse bone marrow before (red line) and after lineage cell depletion (blue line) (JPG, 102 KB)
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Shown is the analysis of bone marrow from one mouse.
- Figure S12. Human HSC progeny in spleen and lung (JPG, 215 KB)
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Human CD34+ cells were transduced with a GFP-expressing lentivirus vectors and transplanted into sublethally irradiated SB17 SCID/beige mice. Nine weeks later, tissue sections were analyzed for GFP by immunohistochemistry using an anti-GFP antibody. GFP-positive cells are brown.
- Figure S13. Rlx expression from MMC cells improves tumor transduction with an adenovirus vector (JPG, 195 KB)
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Neu-tg mice received a s.c. injection of MMC or MMC-Rlx cells. Ten days later Dox was added to drinking water. Fourteen days after tumor cell transplantation, 1 × 108 pfu of an Ad vector expressing b-galactosidase (AdbGal) was injected intratumorally. Tumors were analyzed at day 3 after AdbGal injection for b-gal activity (left panel) and X-gal staining (right panel). N=5. MMC+Dox+AdbGal vs. MMC-Rlx+Dox+Ad.bGal: p=0.008.
- Figure S14 Analysis of tumor-infiltrating cells (JPG, 807 KB)
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Tumor sections from cancer patients were stained for CD68 (brown) and counterstained with hematoxilin. The size bar is 100mm.
- Figure S15. Hematopoietic cells in glioma (JPG, 663 KB)
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Freshly obtained brain/glioma specimens were digested with collagenase at 0.5 mg/ml and DNAse at 25 mg/ml at 37°C for 45 minutes. Following incubation with non-specific mouse IgG1 (clone MOPC 21, Sigma) at 50 mg/ml, about 3 × 105 cells were co-stained with APC-conjugated anti-CD45 mAb (vertical axes) in combination with PE-conjugated anti-KDR, anti-CD33, or anti-CD34 mAb (horizontal axes). Notably, significant amounts of hematopoietic cells were absent in a biopsy of non-malignant tissue (patient #1).
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