Proangiogenic cells enhanced persistent and physiologic neovascularization compared with macrophages

Abstract

Proangiogenic cells (PACs) display surface markers and secrete angiogenic factors similar to those used by myelomonocytic cells, but, unlike myelomonocytic cells, PACs enhance neovascularization activity in experimental ischemic diseases. This study was performed to reveal the differential neovascularization activities of PACs compared with those of myelomonocytic cells. We cultured PACs and CD14+-derived macrophages (Macs) for 7 days. Most of the surface markers and cytokines in the two cell types were alike; the exceptions were KDR, β8 integrin, interleukin-8 and monocyte chemotactic protein-1. Unlike Macs, PACs significantly enhanced mesenchymal stem cell (MSC) transmigration. PACs and Macs increased neovascularization activity in an in vitro co-culture of human umbilical vein endothelial cells and MSCs and in an in vivo cotransplantation in Matrigel. However, the use of Macs resulted in inappropriately dilated and leaky vessels, whereas the use of PACs did not. We induced critical hindlimb ischemia in nude mice, and then transplanted PACs, Macs or vehicle into the mice. We obtained laser Doppler perfusion images weekly. At 2 weeks, mice treated with PACs showed significantly enhanced perfusion recovery in contrast to those treated with Macs. After day 7, when cells were depleted using a suicidal gene, viral thymidine kinase, to induce apoptosis of the cells in vivo by ganciclovir administration, we found that the improved perfusion was significantly abrogated in the PAC-treated group, whereas perfusion was not changed in the Mac-treated group. PACs caused an increase in healthy new vessels in in vitro and in vivo models of angiogenesis and enhanced long-term functional neovascularization activity in the hindlimb ischemia model, whereas Macs did not. Nevertheless, the angiogenic potential and long-term functional results for a specific cell type should be validated to confirm effectiveness and safety of the cell type for use in therapeutic angiogenesis procedures.

Upregulated anti-angiogenic miR-424-5p in type 1 diabetes (model of subclinical cardiovascular disease) correlates with endothelial progenitor cells, CXCR1/2 and other parameters of vascular health

“…In the hindlimb ischemia model, PACs cultured from BPBMCs not only secreted several cytokines such as VEGF and IL-8 (miR-424-5p gene targets (Fig. 4 ) involved in vascular health but also induced well-organised perfused vessels when transplanted which showed neovascularization activity [ 21 ].…”

“…Fifteen random high-power fields (hpf, 200X) photographs were taken, and only dual positive cells were counted. Since PACs have been described as displaying strong neovascularization activity, this assay has been used to study neovascularization potential in studied subjects [ 21 ].…”

Tyrosine-Protein Phosphatase Non-receptor Type 9 (PTPN9) Negatively Regulates the Paracrine Vasoprotective Activity of Bone-Marrow Derived Pro-angiogenic Cells: Impact on Vascular Degeneration in Oxygen-Induced Retinopathy

“…After 5 days of culture, non-adherent cells were removed and adherent cells that internalize acetylated-LDL (acLDL) and express hematopoietic (CD34, CD133, or CD117) and endothelial (lectin, CD31, or VEGFR2) markers were defined as PACs. As schematized in Supplementary Figure 1B, this population of cells mobilizes into ischemic tissues where they promote neovascularization through secretion of angiogenic growth factors (Choi et al, 2015;Rose et al, 2015;Yu et al, 2015;Keighron et al, 2018). Eighty-five (85%) percent of isolated cells in culture were positive for both lectin and acLDL, and 78% of cells were CD34 immunoreactive (Figure 1A).…”

“…Many cell types, mostly hematopoietic progenitors (HPCs), display reparative angiogenic properties in ischemic tissues subjected to myocardial infarction, stroke and limb ischemia (Leeper et al, 2010;Choi et al, 2015;Bian et al, 2019). Of these HPCs, bone marrow (BM)-derived proangiogenic cells (PACs), originally described as early outgrowth endothelial progenitor cells (EPCs), have been shown to integrate ischemic tissues and stimulate angiogenesis by secreting angiogenic factors (Choi et al, 2015;Rose et al, 2015;Keighron et al, 2018).…”

“…Many cell types, mostly hematopoietic progenitors (HPCs), display reparative angiogenic properties in ischemic tissues subjected to myocardial infarction, stroke and limb ischemia (Leeper et al, 2010;Choi et al, 2015;Bian et al, 2019). Of these HPCs, bone marrow (BM)-derived proangiogenic cells (PACs), originally described as early outgrowth endothelial progenitor cells (EPCs), have been shown to integrate ischemic tissues and stimulate angiogenesis by secreting angiogenic factors (Choi et al, 2015;Rose et al, 2015;Keighron et al, 2018). Accordingly, systemic or local administration of naïve or reprogrammed PACs or PAC conditioned media, are effective to promote neovascularization in different animal models of hindlimb ischemia (Ranghino et al, 2012;Yu et al, 2015), myocardial infarction (Schuh et al, 2008;Cheng et al, 2013) and ischemic stroke (Fan et al, 2010;Acosta et al, 2019).…”