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Blood, Vol. 113, Issue 21, 5340-5351, May 21, 2009
Revascularization of ischemic limbs after transplantation of human bone marrow cells with high aldehyde dehydrogenase activity
Blood Capoccia et al.
113: 5340
Supplemental materials for: Capoccia et al
Files in this Data Supplement:
- Figure S1. von Willebrand Factor stained (vWF+) vascular structures were present in the ischemic limbs of mice transplanted with ALDHhi cells (JPG, 74.1 KB)
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Representative photomicrographs of vWF-stained vascular EC in the non-ischemic limb (A) and ischemic limb of mice transplanted with PBS (B), human BM ALDHlo cells (C), or human BM ALDHhi cells (D). Arrowheads represent vWF+ murine EC or small vessel structures (brown) cut in cross-section (arrowheads), or cut longtitudnally (arrows). Mice transplanted with ALDHhi cells showed contiguous small vascular structures between the muscle fibers in the ischemic limb.
- Figure S2. Transplantation of CD14+ monocytes did not increase perfusion in ischemic limbs (JPG, 71.2 KB)
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(A, B) Representative LDPI following right femoral artery ligation and tail vein injection of PBS (n=5), or 1–4.5 × 106 CD14+ monocytes (n=4) isolated from human mobilized peripheral blood samples. Numbers in the lower right of each LDPI image indicate perfusion ratio (PR) of the ischemic versus the non-ischemic limb. (C) Summary of perfusion ratios for mice transplanted as indicated in A–B. Transplantation of CD14+ monocytes did not augment perfusion in ischemic limbs compared to PBS-injected controls.
- Figure S3. Human BM ALDHhi cells produce multipotent hematopoietic colonies in vitro and demonstrate NOD/SCID repopulating function in vivo (JPG, 59.5 KB)
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(A) BM-derived ALDHlo and ALDHhi cells cultured in methylcellulose media with hematopoietic growth factors produced erythroid, granulocyte, and macrophage colonies in vitro. (B) Transplantation of ALDHhi cells into the tail vein of NOD/SCID (●) or NOD/SCID B2M null (○) mice produced consistent hematopoietic engraftment 7–8 weeks post-transplantation (n=19). In contrast, transplanted ALDHlo (■, □) cells did not demonstrate NOD/SCID repopulation (n=10). (C) In mice transplanted with BM ALDHhi cells, human CD45+ cells engrafting the BM showed multipotent differentiation into myeloid (CD33, CD14) and B-lymphoid (CD19, CD20) cells.
- Figure S4. Cell surface marker analysis of human BM-derived ALDHloSSCloand ALDHhiSSClo cells after expansion under mesenchymal or endothelial growth conditions (JPG, 105 KB)
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(A, B) After 28 days of culture in Amniomax™ media, both ALDHlo- and ALDHhi-derived cells did not express hematopoietic (CD45−CD14−), primitive progenitor (CD34−CD133−), or endothelial (CD31−CD144−) cell surface markers, and highly expressed typical stromal fibroblast surface markers (CD73+CD90+). (C) After 28 days in EGM-2 media, ALDHhi-derived cells did not express hematopoietic (CD45−CD14−), primitive progenitor (CD34−CD133−), or endothelial (CD31−CD144−) cell surface markers, and expressed typical stromal fibroblast surface markers (CD73+CD90+). (D) HAEC cultured in EGM-2 media, expressed EC surface markers (CD31+CD144+), but did not express hematopoietic (CD45−CD14−), primitive progenitor (CD34−CD133−), or stromal fibroblast (CD73+CD90−) cell surface markers.
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