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Angiogenic properties of human placenta-derived adherent cells and efficacy in hindlimb ischemia  Aleksandar Francki, PhD, Kristen Labazzo, PhD, Shuyang.

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Presentation on theme: "Angiogenic properties of human placenta-derived adherent cells and efficacy in hindlimb ischemia  Aleksandar Francki, PhD, Kristen Labazzo, PhD, Shuyang."— Presentation transcript:

1 Angiogenic properties of human placenta-derived adherent cells and efficacy in hindlimb ischemia 
Aleksandar Francki, PhD, Kristen Labazzo, PhD, Shuyang He, PhD, Ellen Z. Baum, PhD, Stewart E. Abbot, PhD, Uri Herzberg, DVM, PhD, MBA, Wolfgang Hofgartner, MD, DSc, Robert Hariri, MD, PhD Aleksandr Kaplunovsky, Jennifer Paredes, Allan Reduta, Eric Law, Ewa Fik, Sascha Abramson, Vivian R. Albert, Itschak Lamensdorf Aleksandar Francki, PhD, Kristen Labazzo, PhD, Shuyang He, PhD, Ellen Z. Baum, PhD, Stewart E. Abbot, PhD, Uri Herzberg, DVM, PhD, MBA, Wolfgang Hofgartner, MD, DSc, Robert Hariri, MD, PhD Aleksandr Kaplunovsky, Jennifer Paredes, Allan Reduta, Eric Law, Ewa Fik, Sascha Abramson, Vivian R. Albert, Itschak Lamensdorf  Journal of Vascular Surgery  Volume 64, Issue 3, Pages e1 (September 2016) DOI: /j.jvs Copyright © 2016 Society for Vascular Surgery Terms and Conditions

2 Fig 1 A, Effect of placenta-derived adherent cells (PDACs) on human umbilical vein endothelial cell (HUVEC) survival. HUVECs were exposed to PDACs through a Transwell (PDAC:HUVEC, ∼2:1). The initial value of HUVECs (after 6-hour serum starvation in endothelial cell basal medium and before Transwell exposure) was set to 100%. The difference between HUVEC survival in serum-free Dulbecco's modified Eagle medium (SF-DMEM) overnight (∼35%) and in the presence of PDACs (72%) was statistically significant (P < .01; n = 7). B, Dose-dependent effect of PDACs on HUVEC survival (n = 3). The error bars indicate standard deviation. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Stimulation of human umbilical vein endothelial cell (HUVEC) network tube formation by placenta-derived adherent cell (PDAC)-conditioned medium (CM). Representative photomicrographs of PDAC-induced (upper panel) and serum-free Dulbecco's modified Eagle medium (DMEM) control (lower panel) network (tube) formation are shown. The arrows depict representative tubes. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

4 Fig 3 Quantitation of factors secreted by placenta-derived adherent cells (PDACs). Culture supernatants were analyzed as described for the indicated set of secreted factors. Of the factors examined, hepatocyte growth factor (HGF) is the most abundant, followed by monocyte chemotactic protein 1 (MCP-1) and interleukin-6 (IL-6; n = 5). The error bars indicate standard deviation. FGF-2, Fibroblast growth factor 2; G-CSF, granulocyte colony-stimulating factor; IL-8, interleukin-8; PDGF-BB, platelet-derived growth factor BB; VEGF, vascular endothelial growth factor. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

5 Fig 4 Placenta-derived adherent cells (PDACs) promote vascularization. Percentage change of blood vessel (BV) density was measured in the chorioallantoic membrane (CAM) assay. PDAC-induced vascularization of the CAM was statistically indistinguishable from that of the positive control fibroblast growth factor 2 (FGF-2) and significantly greater than the BV density induced by either the vehicle control (P < .01) or the basal medium (P < .01; n = 7). The error bars indicate standard deviation. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

6 Fig 5 A and B, Placenta-derived adherent cell (PDAC) administration increases blood flow in rodent models of hindlimb ischemia (HLI). Blood flow was measured by noncontact laser Doppler at the indicated times after surgery. PDACs were administered at the indicated doses. Blood flow to the undamaged limb is defined as 100%. Representative experiments from duplicate studies are shown. A, Mouse model. The increase in blood flow in all PDAC treatment groups compared with the vehicle control was statistically significant (from day 28 through day 49; P < .01; n = 15). The error bars indicate standard deviation. B, Rat model. At 35 days, the difference in blood flow between PDAC-treated groups and the vehicle control group is statistically significant (P < .001). Nonviable PDAC control was statistically indistinguishable from the vehicle control (n = 12). The error bars indicate standard deviation. C, Representative in vivo real-time images at different time points of four mice injected in the ischemic hindlimb with PDACs expressing the luciferase reporter gene; mouse No. 5 received vehicle. PDACs persisted <8 days in mice. ROI, Region of interest; VEGF, vascular endothelial growth factor. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

7 Fig 6 Placenta-derived adherent cell (PDAC) treatment increases the angioscore in rat ischemic limb. A, Representative angiographs taken at day 35 after surgery. Rats were treated with (A) vehicle or (B) PDACs (4 × 104/kg). The arrows indicate the surgical site. C, Angioscore of rat hindlimb, 35 days after induction of ischemia. Rats were treated with the indicated doses of PDACs or vehicle control; angioscore of the undamaged limb is defined as 100% (n = 5). The error bars indicate standard deviation. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

8 Fig 7 Histology of cross sections of mouse quadriceps muscle. Quadriceps slices from mice (49 days after surgery) were stained with hematoxylin and eosin; representative micrographs (40× magnification) are shown. The pink shapes are muscle fibers; the white borders are connective tissue. A, Nonsurgical limb. B, Surgical limb from animals treated with vehicle control. C, Vascular endothelial growth factor (VEGF) control. D, Placenta-derived adherent cells (PDACs) at 4 × 104/kg dose. Scale bar = 50 μm. The black dots in B likely represent infiltration of immune cells in the vicinity of a blood vessel. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

9 Fig 8 A, Evaluation of capillary density in cross sections of mouse quadriceps muscle. Quadriceps slices from mice (49 days after surgery) were stained with indoxyl-tetrazolium to detect alkaline phosphatase, a marker of newly synthesized capillary endothelial cells. Representative micrographs (40× magnification) are shown for surgical limbs treated with vehicle control (left panel) or placenta-derived adherent cells (PDACs, 4 × 104/kg dose; right panel). The arrows designate representative areas of alkaline phosphatase staining. Scale bar = 100 μm (n = 5). B, Size distribution of small and large blood vessels in mouse limbs 49 days after surgery. PDACs were administered at the indicated doses (n = 5). The error bars indicate standard deviation. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

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