• Left ventricular remodeling
• Myocardial infarction

• Humans
• Ventricular Remodeling/physiology
• Myocardial Infarction/therapy
• Myocardial Infarction/physiopathology
• Stem Cell Transplantation/methods
• Genetic Therapy/methods

• Humans
• Endothelial Progenitor Cells
• Cell- and Tissue-Based Therapy
• Tissue Engineering
• Thrombosis/therapy
• Hemostasis

• Deep venous thrombosis Gene transfection
• Endothelial progenitor cell
• Lentivirus
• Platelet-derived growth factor

• Animals
• Cell Movement/genetics
• Endothelial Progenitor Cells/metabolism
• Female
• Humans
• Platelet-Derived Growth Factor/metabolism
• Rats
• Rats, Sprague-Dawley
• Thrombosis/metabolism
• Venous Thrombosis/therapy

• Angiogenesis
• CD34 cells
• Fluorescence activated cell sorting (FACS)
• Inflammation
• Magnetic beads cell sorting (MACS)
• Myocardial infarction

• Animals
• Antigens, CD34/metabolism
• Cell Separation/methods
• Cicatrix/pathology
• Female
• Fibrosis
• Flow Cytometry
• Humans
• Immunomodulation
• Inflammation/pathology
• Magnetic Phenomena
• Mice, Inbred NOD
• Mice, SCID
• Myocardial Infarction/physiopathology
• Myocardial Infarction/therapy
• Ventricular Remodeling

• P30 GM127211 NIGMS NIH HHS
• R01 HL127682 NHLBI NIH HHS
• R56 HL124266 NHLBI NIH HHS
• R01 HL138488 NHLBI NIH HHS
• P20 GM103527 NIGMS NIH HHS

CD34+ cells are multipotent hematopoietic stem cells also known as endothelial progenitor cells and are useful in regenerative medicine. Naturally, these cells are mobilized from the bone marrow into peripheral circulation in response to ischemic tissue injury. CD34+ cells are known for their high proliferative and differentiation capacities that play a crucial role in the repair process of myocardial damage. They have an important paracrine activity in secreting factors to stimulate vasculogenesis, reduce endothelial cells and cardiomyocytes apoptosis, remodel extracellular matrix and activate additional progenitor cells. Once they migrate to the target site, they enhance angiogenesis, neovascularization and tissue regeneration. Several trials have demonstrated the safety and efficacy of CD34+ cell therapy in different settings, such as peripheral limb ischemia, stroke and cardiovascular disease. Herein, we review the potential utility of CD34+ cell transplantation in acute myocardial infarction, refractory angina and ischemic heart failure.

• Cell therapy
• Endothelial progenitor cells
• Myocardial ischemia
• Refractory angina
• Heart failure
• Coronary microvascular dysfunction

• bioinformatics
• cord blood
• endothelial progenitor cells
• peripheral blood

Objective: The potential therapeutic role of endothelial progenitor cells (EPCs) in ischemic heart disease for myocardial repair and regeneration is subject to intense investigation. The aim of the study was to investigate the proregenerative potential of human endothelial colony-forming cells (huECFCs), a very homogenous and highly proliferative endothelial progenitor cell subpopulation, in a myocardial infarction (MI) model of severe combined immunodeficiency (SCID) mice. Methods: CD34⁺ peripheral blood mononuclear cells were isolated from patient blood samples using immunomagnetic beads. For generating ECFCs, CD34⁺ cells were plated on fibronectin-coated dishes and were expanded by culture in endothelial-specific cell medium. Either huECFCs (5 × 10⁵) or control medium were injected into the peri-infarct region after surgical MI induction in SCID/beige mice. Hemodynamic function was assessed invasively by conductance micromanometry 30 days post-MI. Hearts of sacrificed animals were analyzed by immunohistochemistry to assess cell fate, infarct size, and neovascularization (huECFCs n = 15 vs. control n = 10). Flow-cytometric analysis of enzymatically digested whole heart tissue was used to analyze different subsets of migrated CD34⁺/CD45⁺ peripheral mononuclear cells as well as CD34⁻/CD45⁻ cardiac-resident stem cells two days post-MI (huECFCs n = 10 vs. control n = 6). Results: Transplantation of human ECFCs after MI improved left ventricular (LV) function at day 30 post-MI (LVEF: 30.43 ± 1.20% vs. 22.61 ± 1.73%, p < 0.001; ΔP/ΔT_max 5202.28 ± 316.68 mmHg/s vs. 3896.24 ± 534.95 mmHg/s, p < 0.05) when compared to controls. In addition, a significantly reduced infarct size (50.3 ± 4.5% vs. 66.1 ± 4.3%, p < 0.05) was seen in huECFC treated animals compared to controls. Immunohistochemistry failed to show integration and survival of transplanted cells. However, anti-CD31 immunohistochemistry demonstrated an increased vascular density within the infarct border zone (8.6 ± 0.4 CD31⁺ capillaries per HPF vs. 6.2 ± 0.5 CD31⁺ capillaries per HPF, p < 0.001). Flow cytometry at day two post-MI showed a trend towards increased myocardial homing of CD45⁺/CD34⁺ mononuclear cells (1.1 ± 0.3% vs. 0.7 ± 0.1%, p = 0.2). Interestingly, we detected a significant increase in the population of CD34⁻/CD45⁻/Sca1⁺ cardiac resident stem cells (11.7 ± 1.7% vs. 4.7 ± 1.7%, p < 0.01). In a subgroup analysis no significant differences were seen in the cardioprotective effects of huECFCs derived from diabetic or nondiabetic patients. Conclusions: In a murine model of myocardial infarction in SCID mice, transplantation of huECFCs ameliorated myocardial function by attenuation of adverse post-MI remodeling, presumably through paracrine effects. Cardiac repair is enhanced by increasing myocardial neovascularization and the pool of Sca1⁺ cardiac resident stem cells. The use of huECFCs for treating ischemic heart disease warrants further investigation.

• adult stem cells
• angiogenesis
• cardiac regeneration
• cardiovascular diseases
• myocardial infraction

• Animals
• Cardiotonic Agents/metabolism
• Diabetes Mellitus/physiopathology
• Disease Models, Animal
• Endothelial Cells/metabolism
• Humans
• Male
• Mice
• Mice, SCID
• Myocardial Infarction/physiopathology
• Tissue Donors

• Endothelin converting enzyme-1
• big endothelin-1
• endothelialization
• shear resistance

Glioblastoma (GBM) is a largely fatal and highly angiogenic malignancy with a median patient survival of just over 1 year with radiotherapy (RT). The effects of RT on GBM remain unclear, although increasing evidence suggests that RT-induced alterations in the brain microenvironment affect the recurrence and aggressiveness of GBM. Glioma stem cells (GSCs) in GBM are resistant to conventional therapies, including RT. This study aimed to investigate the effect of radiation on tumor growth and the GSC microenvironment in a mouse model of glioma. To evaluate the growth-inhibitory effects of ionizing radiation on GSCs, tumor volume was measured via anatomical magnetic resonance imaging (MRI) after the intracranial injection of 1 × 10⁴ human patient-derived GSCs (83NS cells), which exhibit marked radioresistance. When a tumor mass of ~5 mm³ was detected in each animal, 10 Gy of cranial irradiation was administered. Tumor progression was observed in the orthotopic xenografted GSC tumor (primary tumor) from a detectable tumor mass (5 mm³) to a lethal tumor mass (78 mm³) in ~7 d in the non-irradiated group. In the RT group, tumor growth was halted for almost 2 weeks after administering 10 Gy cranial irradiation, with tumor growth resuming thereafter and eventually approaching a lethal mass (56 mm³) 21 d after radiation. Radiation therapy yielded good therapeutic effects, with a 2-fold increase in GSC glioma survival; however, tumor relapse after RT resulted in higher mortality for the mice with a smaller tumor volume (p = 0.029) than the non-irradiated tumor-bearing mice. Moreover, tumor regrowth after IR resulted in different phenotypes associated with glioma aggressiveness compared with the non-irradiated mice; the apparent diffusion coefficient by diffusion MRI decreased significantly (p < 0.05, 0 Gy vs. 10 Gy) alongside decreased angiogenesis, abnormal vascular dilatation, and upregulated CD34, VWF, AQP1, and AQP4 expression in the tumor. These findings demonstrate that radiation affects GSCs in GBM, potentially resulting in therapeutic resistance by changing the tumor microenvironment. Thus, the results of this study suggest potential therapeutic targets for overcoming the resistance of GBMs to RT.

• glioblastoma
• glioma stem cell
• radiotherapy
• tumor microenvironment
• tumor vessel

• cell therapy
• cord blood
• endothelial cells
• endothelial progenitor cell
• ex vivo expansion

• Cell therapy
• Critical limb ischemia
• Endothelial progenitor cells
• Ischemic tissue repair
• Refractory angina

• Breast cancer
• Endothelial progenitor cell
• Hepatocellular cancer
• IL6R
• miR-21

Paralysis of one vocal fold leads to glottal gap and vocal fold insufficiency that has significant impact upon a patient's quality of life. Fillers have been tested to perform intracordal injections, but they do not provide perdurable results. Early data suggest that enriching fat grafts with adipose-derived regenerative cells (ADRCs) promote angiogenesis and modulate the immune response, improving graft survival. The aim of this study is to propose ADRC-enriched adipose tissue grafts as effective filler for the paralyzed vocal fold to use it for functional reconstruction of the glottal gap.

This is the first phase I-IIA clinical trial (phase I/IIA clinical trial, unicentric, randomized, controlled, and two parallel groups), to evaluate the safety of a new therapy with ADRC-enriched fat grafting (ADRC: group I) for laryngoplasty after unilateral vocal fold paralysis. Control group patients received centrifuged autologous fat (CAF: group II) grafts. Overall mean age is 52.49 ± 16.60 years. Group I (ADRC): 7 patients (3 males and 4 females), 52.28 ± 20.95 year. Group II (CAF): 7 patients (3 males and 4 females), 52.71 ± 12.59 year.

VHI-10 test showed that preoperative mean score was 24.21 ± 8.28. Postoperative mean score was 6.71 ± 6.75. Preoperative result in group I was 21.14 ± 3.58 and postoperative result was 3.14 ± 3.53. Preoperative result for group II was 27.29 ± 10.66. Postoperative score in group II was 10.29 ± 7.52. Wilcoxon and the Student t-tests showed that the patient's self-perception of posttreatment improvement is larger when ADRCs are used. Comparing pre- and posttreatment voice quality analysis, group I showed a p = 0.053. Group II showed a p = 0.007. There would be no significant differentiation between pre- and posttreatment results. This is true for group II and limited for group I.

This prospective trial demonstrates the safety and efficacy of the treatment of glottal gap defects utilizing ADRC-enriched fat grafts. This trial is registered with NCT02904824.

Autologous endothelial progenitor cell (EPC) therapy is commonly used to stimulate angiogenesis in ischemic repair and wound healing. However, low total numbers and functional deficits of EPCs make autologous EPC therapy ineffective in diabetes. Currently, no known ex vivo culture techniques can expand and/or ameliorate the functional deficits of EPCs for clinical usage. Recently, we showed that a quality‐quantity culture (QQc) system restores the vasculogenic and wound‐healing efficacy of murine diabetic EPCs. To validate these results and elucidate the mechanism in a translational study, we evaluated the efficacy of this QQc system to restore the vasculogenic potential of diabetic human peripheral blood (PB) CD34+ cells. CD34+ cells purified from PB of diabetic and healthy patients were subjected to QQc. Gene expression, vascular regeneration, and expression of cytokines and paracrine mediators were analyzed. Pre‐ or post‐QQc diabetic human PB‐CD34+ cells were transplanted into wounded BALB/c nude mice and streptozotocin‐induced diabetic mice to assess functional efficacy. Post‐QQc diabetic human PB‐CD34+ cell therapy significantly accelerated wound closure, re‐epithelialization, and angiogenesis. The higher therapeutic efficacy of post‐QQc diabetic human PB‐CD34+ cells was attributed to increased differentiation ability of diabetic CD34+ cells, direct vasculogenesis, and enhanced expression of angiogenic factors and wound‐healing genes. Thus, QQc can significantly enhance the therapeutic efficacy of human PB‐CD34+ cells in diabetic wounds, overcoming the inherent limitation of autologous cell therapy in diabetic patients, and could be useful for treatment of not only wounds but also other ischemic diseases. Stem Cells Translational Medicine2018;7:428–438

• Angiogenesis factor
• Cell therapy
• Culture technique
• Diabetes mellitus
• Wound healing

Autologous transplantation of endothelial progenitor cells (EPCs) is a promising therapeutic approach in the treatment of various vascular diseases. We previously reported a two-step culture system for scalable generation of human EPCs derived from cord blood CD34⁺ cells ex vivo. Here, we now apply this culture system to expand and differentiate human and nonhuman primate EPCs from mobilized peripheral blood (PB) CD34⁺ cells for the therapeutic potential of autologous transplantation.

The human and nonhuman primate EPCs from mobilized PB CD34⁺ cells were cultured according to our previously reported system. The generated adherent cells were then characterized by the morphology, surface markers, nitric oxide (NO)/endothelial NO synthase (eNOS) levels and Dil-acetylated low-density lipoprotein (Dil-Ac-LDL) uptake/fluorescein isothiocyanate (FITC)-lectin binding actives. Furthermore, the efficacy and safety studies were performed by autologous transplantation via hepatic portal vein injection in a nonhuman primate model with acute liver sinusoidal endothelial cell injury.

The mobilized PB CD34⁺ cells from both human and nonhuman primate were efficiently expanded and differentiated. Over 2 × 10⁸ adherent cells were generated from 20 mL mobilized primate PB (1.51 × 10⁶ ± 3.39 × 10⁵ CD34⁺ cells) by 36-day culture and more than 80% of the produced cells were identified as EPCs/endothelial cells (ECs). In the autologous transplant model, the injected EPC/ECs from nonhuman primate PB were scattered in the intercellular spaces of hepatocytes at the hepatic tissues 14 days post-transplantation, indicating successful migration and reconstitution in the liver structure as the functional EPCs/ECs.

We successfully applied our previous two-step culture system for the generation of primate EPCs from mobilized PB CD34⁺ cells, evaluated the phenotypes ex vivo, and transplanted autologous EPCs/ECs in a nonhuman primate model. Our study indicates that it may be possible for these ex-vivo high-efficient expanded EPCs to be used in clinical cell therapy.

• Endothelial progenitor cells
• Hepatic sinusoidal endothelium injury
• Mobilized peripheral blood
• Nonhuman primates

• EPCs
• cardiac
• heart
• ischemia
• progenitors
• reperfusion

• Diabetes
• EPC
• Metformin
• NOX
• ROS
• eNOS

• Animals
• Diabetes Mellitus/drug therapy
• Diabetes Mellitus/pathology
• Endothelial Progenitor Cells/drug effects
• Endothelial Progenitor Cells/pathology
• Humans
• Hypoglycemic Agents/pharmacology
• Hypoglycemic Agents/therapeutic use
• Prognosis
• Reactive Oxygen Species/metabolism
• Telomere Shortening

• CSIR

• Scl2-2 coatings
• cell adhesion
• endothelial progenitor cells
• endothelialization
• migration
• phenotype
• vascular grafts

• bench to clinic
• cardiac regeneration
• myocardial infarction

• Editorials
• angiogenesis
• endothelial progenitor cells
• ischemia
• mononuclear cells

• ischemia
• peripheral arterial disease
• progenitor cell
• stem cell
• vasculogenesis

• Animals
• Cell Differentiation
• Cell Proliferation
• Cell Survival
• Clinical Trials as Topic
• Disease Models, Animal
• Endothelial Cells/metabolism
• Endothelial Cells/transplantation
• Humans
• Limb Salvage
• Neovascularization, Physiologic
• Peripheral Arterial Disease/epidemiology
• Peripheral Arterial Disease/metabolism
• Peripheral Arterial Disease/pathology
• Peripheral Arterial Disease/physiopathology
• Peripheral Arterial Disease/surgery
• Recovery of Function
• Regeneration
• Stem Cell Transplantation
• Time Factors
• Translational Research, Biomedical
• Treatment Outcome

• P01 HL108795 NHLBI NIH HHS
• R01 HL077428 NHLBI NIH HHS
• R01 HL080137 NHLBI NIH HHS
• R01 HL095874 NHLBI NIH HHS

• AC133 Antigen
• Animals
• Antigens, CD/metabolism
• Buffers
• Cell Count
• Cell Culture Techniques/methods
• Cell Culture Techniques/standards
• Cell Proliferation
• Cell- and Tissue-Based Therapy/methods
• Cell- and Tissue-Based Therapy/standards
• Cells, Cultured
• Colony-Forming Units Assay/methods
• Colony-Forming Units Assay/standards
• Culture Media, Serum-Free/metabolism
• Disease Models, Animal
• Dose-Response Relationship, Drug
• Echocardiography
• Endothelial Cells/cytology
• Endothelial Cells/metabolism
• Endothelial Cells/transplantation
• Fetal Blood/cytology
• Fetal Blood/metabolism
• Glycoproteins/metabolism
• Humans
• Immunohistochemistry
• Myocardial Contraction
• Myocardial Infarction/metabolism
• Myocardial Infarction/therapy
• Myocytes, Cardiac/cytology
• Myocytes, Cardiac/metabolism
• Myocytes, Cardiac/transplantation
• Neovascularization, Physiologic
• Peptides/metabolism
• Quality Control
• Rats
• Rats, Nude
• Stem Cells/cytology
• Stem Cells/metabolism
• Ventricular Function, Left

• R01 HL057516 NHLBI NIH HHS
• HL57516 NHLBI NIH HHS
• R37 HL053354 NHLBI NIH HHS
• HL53354 NHLBI NIH HHS
• R01 HL053354 NHLBI NIH HHS

The umbilical cord blood derived endothelial progenitor cells (EPCs) contribute to vascular regeneration in experimental models of ischaemia. However, their ability to participate in cardiovascular tissue restoration has not been elucidated yet. We employed a novel coculture system to investigate whether human EPCs have the capacity to integrate into living and ischaemic cardiac tissue, and participate to neovascularization. EPCs were cocultured with either living or ischaemic murine embryonic ventricular slices, in the presence or absence of a pro-angiogenic growth factor cocktail consisting of VEGF, IGF-1, EGF and bFGF. Tracking of EPCs within the cocultures was performed by cell transfection with green fluorescent protein or by immunostaining performed with anti-human vWF, CD31, nuclei and mitochondria antibodies. EPCs generated vascular tube-like structures in direct contact with the living ventricular slices. Furthermore, the pro-angiogenic growth factor cocktail reduced significantly tubes formation. Coculture of EPCs with the living ventricular slices in a transwell system did not lead to vascular tube-like structures formation, demonstrating that the direct contact is necessary and that the soluble factors secreted by the living slices were not sufficient for their induction. No vascular tubes were formed when EPCs were cocultured with ischaemic ventricular slices, even in the presence of the pro-angiogenic cocktail. In conclusion, EPCs form vascular tube-like structures in contact with living cardiac tissue and the direct cell-to-cell interaction is a prerequisite for their induction. Understanding the cardiac niche and micro-environmental interactions that regulate EPCs integration and neovascularization are essential for applying these cells to cardiovascular regeneration.

• crp
• mcrp
• epc
• cardiovascular disease
• gene array
• interferon-alpha

• Antigens, CD34/metabolism
• C-Reactive Protein/pharmacology
• Cell Differentiation/drug effects
• Cell Differentiation/physiology
• Cell Survival/drug effects
• Cell Survival/physiology
• Cells, Cultured
• Endothelium, Vascular/cytology
• Endothelium, Vascular/drug effects
• Endothelium, Vascular/metabolism
• Humans
• In Vitro Techniques
• Interferon-alpha/metabolism
• Lipoproteins, LDL/metabolism
• Phenotype
• Plant Lectins/metabolism
• Protein Isoforms/pharmacology
• Stem Cells/cytology
• Stem Cells/drug effects
• Stem Cells/metabolism

Endothelial progenitor cells (EPCs) were shown to have angiogenic potential contributing to neovascularization. However, a clear definition of mouse EPCs by cell surface markers still remains elusive. We hypothesized that CD34 could be used for identification and isolation of functional EPCs from mouse bone marrow.

CD34⁺ cells, c-Kit⁺/Sca-1⁺/Lin⁻ (KSL) cells, c-Kit⁺/Lin⁻ (KL) cells and Sca-1⁺/Lin⁻ (SL) cells were isolated from mouse bone marrow mononuclear cells (BMMNCs) using fluorescent activated cell sorting. EPC colony forming capacity and differentiation capacity into endothelial lineage were examined in the cells. Although CD34⁺ cells showed the lowest EPC colony forming activity, CD34⁺ cells exhibited under endothelial culture conditions a more adherent phenotype compared with the others, demonstrating the highest mRNA expression levels of endothelial markers vWF, VE-cadherin, and Flk-1. Furthermore, a dramatic increase in immediate recruitment of cells to the myocardium following myocardial infarction and systemic cell injection was observed for CD34⁺ cells comparing with others, which could be explained by the highest mRNA expression levels of key homing-related molecules Integrin β2 and CXCR4 in CD34⁺ cells. Cell retention and incorporation into the vasculature of the ischemic myocardium was also markedly increased in the CD34⁺ cell-injected group, giving a possible explanation for significant reduction in fibrosis area, significant increase in neovascularization and the best cardiac functional recovery in this group in comparison with the others.

These findings suggest that mouse CD34⁺ cells may represent a functional EPC population in bone marrow, which could benefit the investigation of therapeutic EPC biology.

• Animals
• Antigens, CD34/metabolism
• Bone Marrow Cells/cytology
• Bone Marrow Cells/metabolism
• Cells, Cultured
• Echocardiography
• Endothelial Cells/cytology
• Endothelial Cells/metabolism
• Fluorescent Antibody Technique
• Immunohistochemistry
• Male
• Mice
• Mice, Inbred C57BL
• Mice, Transgenic
• Reverse Transcriptase Polymerase Chain Reaction
• Stem Cells/cytology
• Stem Cells/metabolism

Expanded endothelial progenitor cells (eEPC) improve global left ventricular function in experimental myocardial infarction (MI). Erythropoietin beta (EPO) applied together with eEPC may improve regional myocardial function even further by anti-apoptotic and cardioprotective effects. Aim of this study was to evaluate intramyocardial application of eEPCs and EPO as compared to eEPCs or EPO alone in experimental MI.

In vitro experiments revealed that EPO dosed-dependently decreased eEPC and leukocyte apoptosis. Moreover, in the presence of EPO mRNA expression in eEPC of proangiogenic and proinflammatory mediators measured by TaqMan PCR was enhanced. Experimental MI was induced by ligation and reperfusion of the left anterior descending coronary artery of nude rats (n = 8-9). After myocardial transplantation of eEPC and EPO CD68+ leukocyte count and vessel density were enhanced in the border zone of the infarct area. Moreover, apoptosis of transplanted CD31 + TUNEL + eEPC was decreased as compared to transplantation of eEPCs alone. Regional wall motion of the left ventricle was measured using Magnetic Resonance Imaging. After injection of eEPC in the presence of EPO regional wall motion significantly improved as compared to injection of eEPCs or EPO alone.

Intramyocardial transplantation of eEPC in the presence of EPO during experimental MI improves regional wall motion. This was associated with an increased local inflammation, vasculogenesis and survival of the transplanted cells. Local application of EPO in addition to cell therapy may prove beneficial in myocardial remodeling.