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Manuel Mazo,
Salomón Hernández,
Juan José Gavira,
Gloria Abizanda,
Miriam Araña,
Tania López-Martínez,
Cristina Moreno, Juana Merino,
Alba Martino-Rodríguez,
Alicia Uixeira,
José A García de Jalón,
Juan Pastrana,
Diego Martínez-Caro,
Felipe Prosper
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ABSTRACT: AIMS: To determine the long-term effect of transplantation of adipose-derived stromal cells (ADSCs) in a preclinical model of ischemia/reperfusion (I/R).METHODS AND RESULTS: I/R was induced in 28 Goettingen minipigs by 120 minutes coronary artery occlusion followed by reperfusion. Nine days later, surviving animals were allocated to receive trans-endocardial injection of a mean of 213.6±41.78 million green fluorescent protein (GFP)-expressing ADSCs (n=7) or culture medium as control (n=9). Heart function, cell engraftment and histological analysis were performed 3 months after transplantation. Transplantation of ADSCs induced a statistically significant long-lasting (3 months) improvement in cardiac function and geometry in comparison with control animals.Functional improvement was associated with an increase in angiogenesis and vasculogenesis and a positive effect on heart remodeling with a decrease in fibrosis and cardiac hypertrophy in animals treated with ADSCs. Despite the lack of cell engraftment after 3 months, ADSC transplantation induced changes in the ratio between MMP/TIMP.CONCLUSION: Our results indicate that transplantation of ADSCs, despite the lack of longterm significant cell engraftment, increases vessel density and prevents adverse remodeling in a clinically relevant model of myocardial infarction, strongly suggesting a paracrine mediated effect. ADSCs thus constitute an attractive candidate for the treatment of myocardial infarction.
Cell Transplantation 04/2012; · 5.13 Impact Factor
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ABSTRACT: Anti-human leukocyte antigen (HLA) antibodies are a major cause of allograft loss. Solid-phase immunoassays, notably Luminex technology, have lately begun to replace traditional techniques for detecting these antibodies. This platform, however, carries some restrictions in the type of antibodies it detects. For this reason, results using these new technologies must be correlated with results using traditional techniques that have proven clinical significance. We have correlated flow cytometry cross-match (FCXM) outcomes with results from Luminex assays. Serum samples from patients awaiting transplantation who had known anti-HLA antibodies as detected by Luminex were incubated with lymphocytes expressing (a) 1 of the HLA antigens detected by the sera or (b) several of them. Of the 169 T-cell FCXMs we performed, in 92 cases the target cell expressed only 1 of the HLA antigens detected by the serum. The results obtained correlated well with Luminex data (r = 0.84). A cutoff mean fluorescence intensity value of 6,500 for the Luminex single antigen assay yielded a sensitivity of 85% and specificity of 82% for detecting a positive FCXM. In the other 77 cases, the target cell expressed 2 or more of the HLA antigens detected by the serum. In this situation, the same cutoff proved a useful tool for differentiating negative from positive FCXMs.
Human immunology 03/2012; 73(5):517-21. · 2.55 Impact Factor
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Manuel Mazo,
Arantxa Cemborain,
Juan José Gavira,
Gloria Abizanda,
Miriam Araña,
Mayte Casado,
Mario Soriano,
Salomón Hernández,
Cristina Moreno,
Margarita Ecay,
Edurne Albiasu,
Miriam Belzunce,
Josune Orbe,
José Antonio Páramo, Juana Merino,
Iván Peñuelas,
José Manuel García Verdugo,
Beatriz Pelacho,
Felipe Prosper
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ABSTRACT: Fresh adipose-derived cells have been shown to be effective in the treatment of acute myocardial infarction (MI), but their role in the chronic setting is unknown. We sought to determine the long-term effect of the adipose derived-stromal vascular fraction (SVF) cell transplantation in a rat model of chronic MI. MI was induced in 82 rats by permanent coronary artery ligation and 5 weeks later rats were allocated to receive an intramyocardial injection of 10(7) GFP-expressing fresh SVF cells or culture media as control. Heart function and tissue metabolism were determined by echocardiography and (18)F-FDG-microPET, respectively, and histological studies were performed for up to 3 months after transplantation. SVF induced a statistically significant long-lasting (3 months) improvement in cardiac function and tissue metabolism that was associated with increased revascularization and positive heart remodeling, with a significantly smaller infarct size, thicker infarct wall, lower scar fibrosis, and lower cardiac hypertrophy. Importantly, injected cells engrafted and were detected in the treated hearts for at least 3 months, directly contributing to the vasculature and myofibroblasts and at negligible levels to cardiomyocytes. Furthermore, SVF release of angiogenic (VEGF and HGF) and proinflammatory (MCP-1) cytokines, as well as TIMP1 and TIMP4, was demonstrated in vitro and in vivo, strongly suggesting that they have a trophic effect. These results show the potential of SVF to contribute to the regeneration of ischemic tissue and to provide a long-term functional benefit in a rat model of chronic MI, by both direct and indirect mechanisms.
Cell Transplantation 02/2012; 21(5):1023-37. · 5.13 Impact Factor
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Haematologica 11/2009; 95(4):691-2. · 6.42 Impact Factor
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Manuel Mazo,
Juan José Gavira,
Gloria Abizanda,
Cristina Moreno,
Margarita Ecay,
Mario Soriano,
Pablo Aranda,
María Collantes,
Eduardo Alegría, Juana Merino,
Iván Peñuelas,
José Manuel García Verdugo,
Beatriz Pelacho,
Felipe Prósper
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ABSTRACT: The aim of this study is to assess the long-term effect of mesenchymal stem cells (MSC) transplantation in a rat model of chronic myocardial infarction (MI) in comparison with the effect of bone marrow mononuclear cells (BM-MNC) transplant. Five weeks after induction of MI, rats were allocated to receive intramyocardial injection of 10(6) GFP-expressing cells (BM-MNC or MSC) or medium as control. Heart function (echocardiography and (18)F-FDG-microPET) and histological studies were performed 3 months after transplantation and cell fate was analyzed along the experiment (1 and 2 weeks and 1 and 3 months). The main findings of this study were that both BM-derived populations, BM-MNC and MSC, induced a long-lasting (3 months) improvement in LVEF (BM-MNC: 26.61 +/- 2.01% to 46.61 +/- 3.7%, p < 0.05; MSC: 27.5 +/- 1.28% to 38.8 +/- 3.2%, p < 0.05) but remarkably, only MSC improved tissue metabolism quantified by (18)F-FDG uptake (71.15 +/- 1.27 to 76.31 +/- 1.11, p < 0.01), which was thereby associated with a smaller infarct size and scar collagen content and also with a higher revascularization degree. Altogether, results show that MSC provides a long-term superior benefit than whole BM-MNC transplantation in a rat model of chronic MI.
Cell Transplantation 11/2009; 19(3):313-28. · 5.13 Impact Factor
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Maitane Pérez-Ilzarbe,
María Díez-Campelo,
Pablo Aranda,
Soraya Tabera,
Tania Lopez,
Consuelo del Cañizo, Juana Merino,
Cristina Moreno,
Enrique J Andreu,
Felipe Prósper,
José Antonio Pérez-Simón
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ABSTRACT: Mesenchymal stem cells (MSCs) are multipotent stem cells. Based on their properties, several clinical trials have been designed to explore their potential therapeutic effect. Fetal calf serum (FCS, commonly used for in vitro expansion) is an undesirable source of xenogeneic antigens and bears the risk of transmitting contaminations. As an alternative for FCS, platelet lysate (PL) and both autologous and allogeneic human serum have been proposed. The aim of this study is to compare the culture of bone marrow (BM)-derived MSCs in the presence of different serum supplements to determine the effect on cell growth, differentiation potential, and immunologic function.
MSCs from BM of healthy volunteer donors were grown in the presence of 10% FCS supplemented with 1 ng/mL basic fibroblast growth factor (bFGF), 10% human serum supplemented with 1 ng/mL bFGF, 5% PL, and PL 5% supplemented with 1 ng/mL bFGF (PL plus bFGF).
MSCs that expanded in either medium showed a comparable morphology, phenotype, and proliferative and differentiation capacity. While the presence of MSCs in vitro significantly decreased CD3/CD28-mediated T-cell activation, this effect was significantly higher in MSCs cultured with human serum. Production of interferon-gamma was inhibited by cocultured media with MSCs while MSCs also induced a significant inhibition of cell cycle in T cells.
In conclusion, PL or autologous serum could offer an alternative to the use of FCS in MSC expansion for clinical use maintaining the same growing potential, phenotype, immunomodulatory properties, and differentiation potential.
Transfusion 06/2009; 49(9):1901-10. · 3.22 Impact Factor
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Annals of Hematology 08/2008; 88(2):177-8. · 2.62 Impact Factor
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Miguel Barajas,
Federico Franchi,
Carlos Clavel,
Xabier L Aranguren,
M Gabriela Kramer,
Gloria Abizanda, Juana Merino,
Cristina Moreno,
Leire Gárate,
Anunciata Guitart,
Iñigo Narvaiza,
María Gutiérrez-Perez,
Jose-Ignacio Riezu-Boj,
Carmen Berasain,
Jesús Prieto,
Felipe Prósper
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ABSTRACT: The use of stem cells as a vehicle of therapeutic genes is an attractive approach for the development of new antitumoral strategies based on gene therapy. The aim of our study was to assess the potential of bone marrow-derived Multipotent Adult Progenitor Cells (rMAPCs) to differentiate in vitro and in vivo into endothelial cells and to be recruited to areas of tumor vasculogenesis. In vitro, rMAPCs obtained from Buffalo rats differentiated into cells expressing endothelial markers and demonstrated functional endothelial capacity. Intravenous injection of undifferentiated rMAPC transduced with a lentivirus expressing GFP in an orthotopic rat model of hepatocellular carcinoma, resulted in tumor recruitment of the injected cells and in vivo differentiation into endothelial cells in the tumor area with contribution to vasculogenesis. In summary, our results suggest that rMAPCs can be efficiently recruited by vascularized tumors and differentiate to endothelium and thus may represent a useful vehicle for delivery of therapeutic genes to sites of active tumor neovascularization.
Biochemical and Biophysical Research Communications 01/2008; 364(1):92-9. · 2.48 Impact Factor
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Ricardo García-Muñoz,
Alicia Galar,
Cristina Moreno,
Paula Rodríguez-Otero,
Elena Panizo-Morgado,
Mariano Ponz-Sarvisê,
Mirian Fernandez-Alonso,
Manuel Rubio, Juana Merino,
Braulia Cuesta,
Carlos Panizol,
Felipe Prosper
Leukemia and Lymphoma 01/2008; 48(12):2461-4. · 2.58 Impact Factor
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Xabier L Aranguren,
Aernout Luttun,
Carlos Clavel,
Cristina Moreno,
Gloria Abizanda,
Miguel A Barajas,
Beatriz Pelacho,
Maialen Uriz,
Miriam Araña,
Ana Echavarri,
Mario Soriano,
Enrique J Andreu, Juana Merino,
Jose Manuel Garcia-Verdugo,
Catherine M Verfaillie,
Felipe Prósper
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ABSTRACT: Many stem cell types have been shown to differentiate into endothelial cells (ECs); however, their specification to arterial or venous endothelium remains unexplored. We tested whether a specific arterial or venous EC fate could be induced in human multipotent adult progenitor cells (hMAPCs) and AC133(+) cells (hAC133(+)). In vitro, in the presence of VEGF(165), hAC133(+) cells only adopted a venous and microvascular EC phenotype, while hMAPCs differentiated into both arterial and venous ECs, possibly because hMAPCs expressed significantly more sonic hedgehog (Shh) and its receptors as well as Notch 1 and 3 receptors and some of their ligands. Accordingly, blocking either of those pathways attenuated in vitro arterial EC differentiation from hMAPCs. Complementarily, stimulating these pathways by addition of Delta-like 4 (Dll-4), a Notch ligand, and Shh to VEGF(165) further boosted arterial differentiation in hMAPCs both in vitro and in an in vivo Matrigel model. These results represent the first demonstration of adult stem cells with the potential to be differentiated into different types of ECs in vitro and in vivo and provide a useful human model to study arteriovenous specification.
Blood 04/2007; 109(6):2634-42. · 9.90 Impact Factor
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ABSTRACT: Toll-like receptors recognize specific patterns of microbial components and regulate the activation of both innate and adaptive immunity. TLR4 recognizes lipopolysaccharide (LPS) in monocytes/macrophages with the help of other molecules like CD14 and MD-2, which indicates that the functional LPS receptor forms a large complex. The functional relationship between the components has been the subject of debate, as have the modifications induced by the ligand in the expression of some of these components. Moreover, as for other members of this family of receptors, the possible direct interaction of receptors and their ligands is a matter of discussion. In this paper we address the question of whether the expression of some of the components influences the expression of the rest. Human monocytes in which CD14 has been downregulated through interference in the turnover of the molecule at the Golgi level, show normal membrane TLR4 expression, when compared with control cells. On the other hand, LPS alters membrane TLR4 expression by monocytes devoid of membrane CD14 only in the presence of human serum. The effect of serum is blocked by anti-CD14 monoclonal antibodies, which strongly suggests a functional role for soluble CD14/LPS complexes in the interaction with TLR4. Our data add information on the relationship between the components of the LPS receptor and the characteristics of the interaction of LPS and TLR4 in cells devoid of membrane CD14.
Microbes and Infection 10/2004; 6(11):990-5. · 3.10 Impact Factor
