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1.
Stem Cell Res ; 17(1): 6-15, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27186654

RESUMO

Successful stem cell therapy after acute myocardial infarction (AMI) is hindered by lack of engraftment of sufficient stem cells at the site of injury. We designed a novel technique to overcome this problem by assembling stem cell-microbubble complexes, named 'StemBells'. StemBells were assembled through binding of dual-targeted microbubbles (~3µm) to adipose-derived stem cells (ASCs) via a CD90 antibody. StemBells were targeted to the infarct area via an ICAM-1 antibody on the microbubbles. StemBells were characterized microscopically and by flow cytometry. The effect of ultrasound on directing StemBells towards the vessel wall was demonstrated in an in vitro flow model. In a rat AMI-reperfusion model, StemBells or ASCs were injected one week post-infarction. A pilot study demonstrated feasibility of intravenous StemBell injection, resulting in localization in ICAM-1-positive infarct area three hours post-injection. In a functional study five weeks after injection of StemBells cardiac function was significantly improved compared with controls, as monitored by 2D-echocardiography. This functional improvement neither coincided with a reduction in infarct size as determined by histochemical analysis, nor with a change in anti- and pro-inflammatory macrophages. In conclusion, the StemBell technique is a novel and feasible method, able to improve cardiac function post-AMI in rats.


Assuntos
Microbolhas , Infarto do Miocárdio/terapia , Transplante de Células-Tronco/métodos , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Administração Intravenosa , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Sobrevivência Celular , Células Cultivadas , Modelos Animais de Doenças , Ecocardiografia , Coração/diagnóstico por imagem , Coração/fisiopatologia , Humanos , Macrófagos/metabolismo , Macrófagos/patologia , Masculino , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Projetos Piloto , Ratos , Ratos Wistar , Sonicação , Células-Tronco/citologia , Células-Tronco/metabolismo
2.
Cell Tissue Res ; 362(3): 623-32, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26202892

RESUMO

In most pre-clinical animal studies investigating stem cell therapy in acute myocardial infarction (AMI), the administered stem cells are isolated from healthy donors. In clinical practice, however, patients who suffer from AMI will receive autologous cells, for example using adipose-derived stem cells (ASC). During AMI, inflammation is induced and we hypothesized that this might affect characteristics of ASC. To investigate this, ASC were isolated from rat adipose tissue 1 day (1D group, n = 5) or 7 days (7D group, n = 6) post-AMI, and were compared with ASC from healthy control rats (Control group, n = 6) and sham-operated rats (Sham 1D group, n = 5). We found that significantly fewer ASC were present 1 day post-AMI in the stromal vascular fraction (SVF), determined by a colony-forming-unit assay (p < 0.001 vs. Control and 7D). These data were confirmed by flow cytometry, showing fewer CD90-positive cells in SVF of the 1D group. When cultured, no differences were found in proliferation rate and cell size between the groups in the first three passages. Also, no difference in the differentiation capacity of ASC was found. In conclusion, it was shown that significantly fewer stem cells were present in the SVF 1 day post-AMI; however, the stem cells that were present showed no functional differences.


Assuntos
Tecido Adiposo/citologia , Infarto do Miocárdio/patologia , Células-Tronco/citologia , Animais , Contagem de Células , Diferenciação Celular , Linhagem da Célula , Células Cultivadas , Masculino , Ratos Wistar , Células Estromais/citologia
3.
Stem Cell Res ; 13(3 Pt A): 367-78, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25290189

RESUMO

Adipose tissue-derived stromal cells (ASC) form a rich source of autologous cells for use in regenerative medicine. In vitro induction of an endothelial phenotype may improve performance of ASCs in cardiovascular repair. Here, we report on an in vitro strategy using direct reprogramming of ASCs by means of ectopic expression of the endothelial-specific transcription factor SRY (sex determining region Y)-box18 (SOX18). SOX18 induces ASCs to express a set of genes involved in vascular patterning: MMP7, KDR, EFNB2, SEMA3G and CXCR4. Accordingly, SOX18 transduced ASCs reorganize under conditions of shear stress, display VEGF-induced chemotaxis and form tubular structures in 3D matrices in an MMP7-dependent manner. These in vitro findings provide insight into molecular and cellular processes downstream of SOX18 and show that reprogramming using SOX18 is sufficient to induce several endothelial-like features in ASCs.


Assuntos
Tecido Adiposo/citologia , Células Endoteliais/metabolismo , Fatores de Transcrição SOXF/metabolismo , Células Estromais/metabolismo , Diferenciação Celular , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Reprogramação Celular , Quimiotaxia/efeitos dos fármacos , Células Endoteliais/citologia , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Metaloproteinase 7 da Matriz/metabolismo , Microtúbulos/química , Microtúbulos/metabolismo , Fatores de Transcrição SOXF/genética , Resistência ao Cisalhamento , Células Estromais/citologia , Fator A de Crescimento do Endotélio Vascular/farmacologia
4.
Res Vet Sci ; 96(2): 377-9, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24445251

RESUMO

The Wistar rat is a commonly used strain for experimental animal models. Recently it was shown that results vary between studies using Wistar rats of different suppliers. Therefore we studied whether Wistar rats obtained from Harlan Laboratories (Ha, n=24) and Charles River (CR, n=22) had a different outcome in an acute myocardial infarction (AMI) model. AMI was induced in both Ha and CR Wistar rats by one operator. This resulted in a significantly higher survival rate for Ha (79.2±10.2%) compared with CR rats (54.2±10.2%, p<0.05). Furthermore, CR rats had lost significantly more weight after 7 days (-5.9±3.1%) compared with Ha rats (-0.8±1.7%; p<0.001), indicating a worse health status of the CR rats. Paradoxically, the induced infarct was smaller in CR rats (7.3±3.6% of the heart) compared with Ha rats (12.1±4.7%, p<0.05). This indicates that CR rats were less sensitive for the cardiomyocyte damage subsequent to AMI induction, but remarkably showed more clinical side effects indicating that Wistar rats from two suppliers had a different response within the same AMI model.


Assuntos
Infarto do Miocárdio/veterinária , Miócitos Cardíacos/ultraestrutura , Ratos Wistar/cirurgia , Animais , Modelos Animais de Doenças , Histocitoquímica , Estimativa de Kaplan-Meier , Masculino , Infarto do Miocárdio/fisiopatologia , Ratos
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