Placenta-derived PLX-PAD mesenchymal-like stromal cells are efficacious in rescuing blood flow in hind limb ischemia mouse model by a dose- and site-dependent mechanism of action

Abstract Backgroud: Human umbilical cord blood (UCB) contains a high number of endothelial progenitor cells (EPCs) and may be useful for the treatment of ischemic disease. Recently, we have isolated EPCs from UCB having different biologic properties for angiogenic capabilities in vitro. In this present study, the aim is to examine the usefulness of OECs in hind-limb ischemia. Methods: Mononuclear cells from UCB cultured using EGM-2 medium with VEGF, IGF-1 and FGF for 21 days. Early spindle-shaped cells (early OECs), which were grown during the first week of culture and late cobblestone shaped cells (late OECs), which were in peak growth during the third week of culture were found. The hind-limb ischemia was established as follows: Athymic nude mice (BALB/C-nu) 18–22 g in weight were anesthetized with pentobarbital (60 mg/kg) and their left femoral arteries and main extension arteries were operatively resected. To examine the effect of the vasculogenesis of the two types of OECs, the mice were divided into three groups (PBS, early and late OECs). Twenty-four hours after operative excision 5 × 105 OECs in 200 μl and an equal volume of PBS were administered by intramuscular injection into the mice on hind-limb ischemia. To compare the effect of OECs on neovascularization in vivo, the analysis of blood flow of ischemic and healthy hind limbs was performed on days 1 and 21 after surgery using near-infrared (NIR) imaging with incocyanne Green (ICG). Results: Late OECs expressed a high level of mRNA on endothelial marker genes and formed capillary tubes in Matrigel plates. The early spindle cells excreted more angiogenic cytokines and had more migratory ability. We divided the mice into two groups according to the degree of perfusion; good (22.5–50%/min) and poor (0–22.5%) perfusion. OECs improved the blood flow of the ischemic hind-limb in the ’good’ perfusion group but not in the ’poor’ perfusion group. Early OECs led to a more significant improvement in blood flow than that of the late OECs. Conclusion: The different types of OECs from UCB have different biologic properties in vitro and different vasculogenic potential in vivo as well. The results might have potential application for the treatment of hind-limb ischemia.

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SS5. SVS Foundation Resident Research Prize Winner A Novel Mouse Model of Hind Limb Ischemia to Test Therapeutic Angiogenesis

Journal of Vascular Surgery ◽

10.1016/j.jvs.2012.03.071 ◽

2012 ◽

Vol 55 (6) ◽

pp. 18S-19S

Author(s):

Robert Brenes ◽

Caroline C. Jadlowiec ◽

Mackenzie Bear ◽

Peter Hashim ◽

Clinton D. Protack ◽

...

Keyword(s):

Mouse Model ◽

Hind Limb ◽

Limb Ischemia ◽

Therapeutic Angiogenesis ◽

Hind Limb Ischemia ◽

Resident Research

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Effects on duration of post-operative ischemia and patterns of blood flow recovery in different conditions of mouse hind limb ischemia

Vascular Cell ◽

10.1186/2045-824x-3-14 ◽

2011 ◽

Vol 3 (1) ◽

pp. 14 ◽

Cited By ~ 1

Author(s):

Husain A Al-Mubarak ◽

Talal M Alamri ◽

Saif A Aljabab ◽

Mohammad Atteya ◽

Adrian Quan ◽

...

Keyword(s):

Blood Flow ◽

Hind Limb ◽

Limb Ischemia ◽

Hind Limb Ischemia

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Abstract 456: Rage Suppresses Angiogenesis and Blood Flow Recovery After Hind Limb Ischemia in Diabetic Mice Through Modulation of Macrophage Inflammation and Macrophage-endothelial Interactions

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.37.suppl_1.456 ◽

2017 ◽

Vol 37 (suppl_1) ◽

Author(s):

Raquel Lopez Diez ◽

Qing Li ◽

Huilin Li ◽

Shi Fang Yan ◽

Ann Marie Schmidt

Keyword(s):

Blood Flow ◽

Hind Limb ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Limb Ischemia ◽

Diabetic Mice ◽

Hind Limb Ischemia ◽

Inflammatory Monocytes ◽

Baseline Levels ◽

Ischemic Muscle

Peripheral vascular disease is a condition characterized by atherosclerotic narrowed arteries distal to the aorta which triggers an acute or critical limb ischemia. Development of ischemic PVD has been considered one of the principal complications of diabetes, leading to amputation of digits and limbs. Advanced glycation end products (AGE) ligands and their receptor (RAGE) have been implicated in multiple key mechanisms underlying diabetes and diabetic complications, including hypoxia and ischemia/reperfusion injury. We tested the hypothesis that vascular recovery after hind limb ischemia would be rescued by deficiency of RAGE, at least in part through modulation of macrophage dysfunction. Wild type (WT) and Ager deficient mice were rendered diabetic with streptozotocin, and subjected to unilateral hind limb ischemia. Previous results showed an increased accumulation and expression of AGEs and RAGE in ischemic muscle, especially in diabetic WT mice. Attenuated angiogenesis and impaired blood flow recovery were also observed, in parallel with reduced early inflammatory macrophage infiltration into ischemic muscle in the WT diabetic mice. We performed flow cytometry to analyze circulating monocyte subsets: pro-inflammatory Ly6G/C hi and anti-inflammatory Ly6G/C lo . Work by others reported higher levels of monocytes in diabetes in the baseline state without injury; our data indicate that the increase is mostly attributed to the pro-inflammatory Ly6G/C hi population, being significantly lower in the Ager -/- diabetic mice. After seven days of ischemia to the unilateral hind limb, lower levels of circulating pro-inflammatory Ly6G/C hi monocytes were found in both WT and Ager deficient diabetic mice. After four weeks of injury, the pro-inflammatory monocytes levels were significantly recovered to baseline levels in Ager -/- mice, whereas WT mice failed to reacquire their baseline levels. In vitro studies using murine endothelial cells and murine macrophages revealed that RAGE suppressed macrophage-endothelial cell interaction, particularly in diabetes-relevant concentrations of D-glucose. These data suggest unique ischemia-dependent mechanisms in hind limb ischemia through RAGE down-regulation of the early adaptive immune response.

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