Human Peripheral Blood-Derived CD31+Cells Have Robust Angiogenic and Vasculogenic Properties and Are Effective for Treating Ischemic Vascular Disease

Abstract

Objectives This study aimed to determine if CD31 is a novel marker of a circulating angio-vasculogenic cell population and to establish their therapeutic effects on experimental ischemia. Background Emerging evidence suggested that therapeutic mechanisms underlying various bone marrow (BM)-derived cells are due to paracrine effects. Furthermore, the vasculogenic potential of these cells is under debate. CD31 is a well known marker for endothelial cells (ECs) but is also expressed in a fraction of peripheral blood (PB) mononuclear cells. Methods CD31+ cells were isolated from human PB by magnetic-activated cell sorting (MACS). The gene expression profile was examined by DNA microarray and real-time RT-PCR (qRT-PCR). Various in vitro endothelial differentiation or vasculogenic assays were conducted. Finally, cells were directly implanted into a mouse hindlimb ischemia (HLI) model to test angiogenic-vasculogenic and therapeutic effects. Results Fluorescent-activated cell sorter (FACS) analysis revealed that PB-CD31+ cells exhibited endothelial and hematopoietic stem/progenitor markers. CD31+ cells had higher levels of expression of pro-angiogenic genes on microarray and qRT-PCR and generated higher numbers of endothelial progenitor cells (EPCs) compared to CD31− cells. CD31+ cells spontaneously formed vascular tube-like structures and exhibited an endothelial cell phenotype in vitro. In a HLI model, CD31+ cell transplantation augmented blood perfusion and prevented limb loss. Both angiogenic cytokines and capillary density were increased, suggesting CD31+ cells augmented neovascularization. Conclusions CD31 is a novel marker that designates circulating angiogenic and vasculogenic cells. These cells are easily isolated from human PB and thus are a novel candidate for treatment of ischemic cardiovascular disease.

A stromal cell-derived factor-1 releasing matrix enhances the progenitor cell response and blood vessel growth in ischaemic skeletal muscle

“…By one week post-treatment, animals that received matrix treatment (with or without SDF-1 microspheres) had hindlimb perfusion restored to baseline levels, which is similar to previous reports of matrix treatment for hindlimb ischaemia . It has been previously demonstrated that an increase in perfusion is attributable to increased local vasculature (Kim et al, 2010;Suuronen et al, 2010), an observation that was seen in the current study; matrix treatments increased arteriole density in ischaemic hindlimbs. It was hypothesised that SDF-1 microsphere matrix treatment would confer superior restoration of perfusion, but instead, the matrix-only treatment was equally effective.…”

Angio-Vasculogenic Properties of Endothelial-Induced Mesenchymal Stem Cells Derived From Human Adipose Tissue

“…Animal experiments were performed as described previously. 12 Mice were anesthetized with isoflurane (induction: 450 ml air, 4.5% isoflurane; maintenance: 200 ml air, 2.0% isoflurane; Baxter International, Inc, Deerfield, IL, USA), and the depth of anesthesia was monitored by respiratory rate and lack of withdrawal reflex upon toe pinching. 4 Hindlimb ischemia was induced by ligation of the right femoral artery.…”

Coxsackievirus B3 used as a gene therapy vector to express functional FGF2

“…21 --23 Therefore, we generated ischemic hindlimbs by unilateral femoral artery ligation, as previously described, 24,25 and then injected them with CVB3 --FGF2, CVB3 --GFP, CVB3 or PBS. CVB3 --FGF2 significantly protected the hindlimbs from ischemic necrosis compared with PBS, CVB3 and CVB3 --GFP in BALB/c mice (Po0.05; Figure 4), whereas there were no differences in limb salvage among the PBS, CVB3 and CVB3 --GFP groups, which all showed significant limb necrosis (Figure 4).…”