Name: Análise abrangente de plaquetas pró-coagulantes exibindo características de necrose, apoptose e ativação plaquetária (Vídeo) | JoVE Journal
Uploaded: 2025-05-23T14:00:00.000+00:00
Duration: 4 min 37 s
Description: Aprenda a analisar plaquetas pró-coagulantes com características de necrose, apoptose e ativação em plaquetas humanas lavadas. Este método auxilia na identificação de biomarcadores para trombose e alvos terapêuticos.

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<li>Aliotta, A., Bertaggia Calderara, D., Zermatten, M. G., Marchetti, M., Alberio, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Thrombocytopathies%3A+Not+just+aggregation+defects-the+clinical+relevance+of+procoagulant+platelets%5BTitle%5D)+AND+%22J+Clin+Med%22%5BJournal%5D)">Thrombocytopathies: Not just aggregation defects-the clinical relevance of procoagulant platelets.</a> J Clin Med. 10 (5), 894(2021).</li>
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<li>Kholmukhamedov, A., Janecke, R., Choo, H. -J., Jobe, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+mitochondrial+calcium+uniporter+regulates+procoagulant+platelet+formation%5BTitle%5D)+AND+%22J+Thromb+Haemost%22%5BJournal%5D)">The mitochondrial calcium uniporter regulates procoagulant platelet formation.</a> J Thromb Haemost. 16 (11), 2315-2321 (2018).</li>
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<li>Schoenwaelder, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Two+distinct+pathways+regulate+platelet+phosphatidylserine+exposure+and+procoagulant+function%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Two distinct pathways regulate platelet phosphatidylserine exposure and procoagulant function.</a> Blood. 114 (3), 663-666 (2009).</li>
<li>Abbasian, N., Millington-Burgess, S. L., Chabra, S., Malcor, J. -D., Harper, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Supramaximal+calcium+signaling+triggers+procoagulant+platelet+formation%5BTitle%5D)+AND+%22Blood+Adv%22%5BJournal%5D)">Supramaximal calcium signaling triggers procoagulant platelet formation.</a> Blood Adv. 4 (1), 154-164 (2020).</li>
<li>Le, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+inner+activation+gate+controls+TMEM16F+phospholipid+scrambling%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">An inner activation gate controls TMEM16F phospholipid scrambling.</a> Nat Commun. 10 (1), 1846(2019).</li>
<li>Yang, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((TMEM16F+forms+a+Ca2%2B-activated+cation+channel+required+for+lipid+scrambling+in+platelets+during+blood+coagulation%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">TMEM16F forms a Ca2+-activated cation channel required for lipid scrambling in platelets during blood coagulation.</a> Cell. 151 (1), 111-122 (2012).</li>
<li>Mattheij, N. J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dual+mechanism+of+integrin+closure+in+procoagulant+platelets%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Dual mechanism of integrin closure in procoagulant platelets.</a> J Biol Chem. 288 (19), 13325-13336 (2013).</li>
<li>Liu, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrially+mediated+integrin+%CE%B1iIb%CE%B23+protein+inactivation+limits+thrombus+growth%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Mitochondrially mediated integrin αiIbβ3 protein inactivation limits thrombus growth.</a> J Biol Chem. 288 (42), 30672-30681 (2013).</li>
<li>van Kruchten, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Both+TMEM16F-dependent+and+TMEM16F-independent+pathways+contribute+to+phosphatidylserine+exposure+in+platelet+apoptosis+and+platelet+activation%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Both TMEM16F-dependent and TMEM16F-independent pathways contribute to phosphatidylserine exposure in platelet apoptosis and platelet activation.</a> Blood. 121 (10), 1850-1857 (2013).</li>
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<li>Shlomovitz, I., Speir, M., Gerlic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Flipping+the+dogma+-+phosphatidylserine+in+non-apoptotic+cell+death%5BTitle%5D)+AND+%22Cell+Commun+Signal%22%5BJournal%5D)">Flipping the dogma - phosphatidylserine in non-apoptotic cell death.</a> Cell Commun Signal. 17 (1), 139(2019).</li>
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<li>Michelson, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Platelet+activation+by+thrombin+can+be+directly+measured+in+whole+blood+through+the+use+of+the+peptide+GPRP+and+flow+cytometry%3A+Methods+and+clinical+applications%5BTitle%5D)+AND+%22Blood+Coagul+Fibrinolysis%22%5BJournal%5D)">Platelet activation by thrombin can be directly measured in whole blood through the use of the peptide GPRP and flow cytometry: Methods and clinical applications.</a> Blood Coagul Fibrinolysis. 5 (1), 121-131 (1994).</li>
<li>Taylor, M. R., Couto, J. R., Scallan, C. D., Ceriani, R. L., Peterson, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lactadherin+%28formerly+BA46%29%2C+a+membrane-associated+glycoprotein+expressed+in+human+milk+and+breast+carcinomas%2C+promotes+Arg-Gly-Asp+%28RGD%29-dependent+cell+adhesion%5BTitle%5D)+AND+%22DNA+Cell+Biol%22%5BJournal%5D)">Lactadherin (formerly BA46), a membrane-associated glycoprotein expressed in human milk and breast carcinomas, promotes Arg-Gly-Asp (RGD)-dependent cell adhesion.</a> DNA Cell Biol. 16 (7), 861-869 (1997).</li>
<li>Andersen, M. H., Graversen, H., Fedosov, S. N., Petersen, T. E., Rasmussen, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Functional+analyses+of+two+cellular+binding+domains+of+bovine+lactadherin%5BTitle%5D)+AND+%22Biochemistry%22%5BJournal%5D)">Functional analyses of two cellular binding domains of bovine lactadherin.</a> Biochemistry. 39 (20), 6200-6206 (2000).</li>
<li>Tan, C. W., Bourcy, M., Pasalic, L., Chen, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Flow+cytometry+assessment+of+procoagulant+platelets+using+a+dithiol-reactive+probe+functional+disulfide+bonds%3A+Methods+and+protocols%5BTitle%5D)+AND+%22Methods+Mol+Biol%22%5BJournal%5D)">Flow cytometry assessment of procoagulant platelets using a dithiol-reactive probe functional disulfide bonds: Methods and protocols.</a> Methods Mol Biol. 1967, 305-321 (2019).</li>
</ol>

<table><tbody><tr><td>Solução de Citrato Ácido Dextrose (ACD) (Para 1000 mL)</td><td></td><td></td><td>Citrato de Tri-Sódio- &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 22 g Ácido Cítrico-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8 g Dextrose-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 24,5 g Água-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Volume de reposição para 1000 mL&nbsp;</td></tr><tr><td>Kit de rotulagem de proteínas Alexa Fluor 488</td><td>Invitrogen</td><td>A10235</td><td></td></tr><tr><td>APC Mouse Anti-Human CD62P</td><td>BD Pharmingen</td><td>550888</td><td></td></tr><tr><td>Protrombina Bovina</td><td>Prolytix</td><td>BCP-1010</td><td></td></tr><tr><td>Buffer A (Preparação de Plaquetas)</td><td></td><td></td><td>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; M.W&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. em 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 100 mL de solução 10X HEPES&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238,30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 20 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.766 g NaCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 58.44&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 134 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7,83 g KCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 74,55&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2,9 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 216,19 mg MgCl2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203,30 mg NaH2PO4&nbsp; &nbsp; &nbsp; 156,01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 0,34 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 53,04 mg NaHCO3&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 84 01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 12 mM &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1,01 g Água para 100 mL após ajustar o pH para 6,2 Diluir a solução 10X 1:10 (v/v) com água Milli Q imediatamente antes da preparação das plaquetas para obter a solução 1X que precisa ser complementada com o seguinte &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. em 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 1 ml de solução 1X EGTA &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Adicione 10 &mu; L (1:100 v/v) 100 mM EGTA&nbsp; Glicose &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 mM &nbsp; &nbsp; &nbsp; &nbsp; Adicione 5 &mu; L (1:200 v/v) 1 M glicose PGE1 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3 &mu; M &nbsp; &nbsp; &nbsp; &nbsp; Adicione 3 &mu; L (1:333 v/v) 1 mM PGE1 solução</td></tr><tr><td>Tampão B (Preparação de Plaquetas)</td><td></td><td></td><td>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; M.W&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. em 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 100 mL de solução 10X HEPES&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238,30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 20 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.766 g NaCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 58.44&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 134 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7,83 g KCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 74,55&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2,9 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 216,19 mg MgCl2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203,30 mg NaH2PO4&nbsp; &nbsp; &nbsp; 156,01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 0,34 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 53,04 mg NaHCO3&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 84 01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 12 mM &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1,01 g Água para 100 mL após ajuste do pH para 7,4 Diluir a solução 10X 1:10 (v/v) com água Milli Q imediatamente antes da preparação das plaquetas para obter a solução 1X que precisa ser complementada com o seguinte &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. em 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 1 ml de solução 1X Glicose &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 mM &nbsp; &nbsp; &nbsp; &nbsp; Adicione 5 &mu; L (1:200 v/v) 1 M glicose&nbsp;</td></tr><tr><td>CaCl2 (0,5 M) (Para 50ml)</td><td></td><td></td><td>Peso molecular de CaCl2.2H2O = 147,02 Dissolver 3,675 g de CaCl2.2H2O em 50 ml de água Milli Q</td></tr><tr><td>CellEvent Caspase-3/7 Reagentes de Detecção Green</td><td>Invitrogen</td><td>C10423</td><td>Reagente de detecção de apoptose</td></tr><tr><td>Convulxin</td><td>Enzo Life Sciences</td><td>ALX-350-100</td><td></td></tr><tr><td>EDTA (0,5 M; pH 8) (Para 100 mL)</td><td></td><td></td><td>Peso Molecular do EDTA Na2.2H2 O: 372,24g Pesar 18,612 g e suspender em 50 ml de água Milli Q e verificar o pH (pH~4) Adicionar lentamente 10N NaOH com agitação e monitorar o pH. O EDTA começa a solubilizar em torno de pH 7 e é completamente solúvel em pH 8, Completar o volume para 100 ml com água Milli Q.</td></tr><tr><td>EGTA (100 mM; pH 7,4) (Para 100 mL)</td><td></td><td></td><td>Peso Molecular: 380,4 Adicionar 3,804 g de EGTA em 50 ml de água Milli Q e verificar o pH (pH~3) Adicionar 10 N NaOH gota a gota enquanto agita e monitorar o pH EGTA torna-se solúvel em pH 7,0 (aprox) Ajustar o pH para 7,4 Completar o volume para 100 ml com água</td></tr> <tr><td>Milli QKit de coloração FITC Mouse Anti-Human PAC-1</td><td>BD</td><td>BD 340507</td><td></td></tr><tr><td>FITC-IETD-FMK Caspase 8 (ativo) Abcam</td><td></td><td>ab65614</td><td></td></tr><tr><td>Mitotracker Red CMXRos (corante de marcação de mitochandria)</td><td>Invitrogen</td><td>M7512</td><td>Estoque= 1 mM (50 &micro; g dissolvido em 90 &micro; l DMSO) Sub-estoque= 100 &micro; M (10 &micro; l Estoque + 90 &micro; l DMSO) Concentração de trabalho= 500nM (0,5 &micro; l em 100 &micro; l)</td></tr> <tr><td>PE Anexina V</td><td>BD Pharmingen</td><td>560930</td><td></td></tr><tr><td>Prostaglandina E1</td><td>Sigma</td><td>P5515</td><td>Estoque = 20 mM (1 mg dissolvido em 141 &micro; L DMSO) Sub-estoque= 1 mM (10 &micro; l Estoque + 190 &micro; L DMSO) Concentração de trabalho= 3 &micro; M (3 &micro; L em 1 mL)</td></tr><tr><td>Rhod-2 AM</td><td>Invitrogen</td><td>R1244</td><td>Stock= 5 mM (1 mg dissolvido em 178 &micro; L DMSO) Sub-estoque= 100 &micro; M (10 &micro; L Stock + 90 &micro; L DMSO) Concentração de trabalho= 500 nM (0,5 &micro; L em 100 &micro; L)</td></tr> <tr><td>Trombina do plasma humano</td><td>Sigma</td><td>T7572</td><td></td></tr></tbody></table>

análise abrangente de plaquetas pró-coagulantes exibindo características de necrose, apoptose e ativação plaquetária

Comprehensive Analysis of Procoagulant Platelets Exhibiting Features of Necrosis, Apoptosis and Platelet Activation

Uma proporção de plaquetas ativadas torna-se "pró-coagulante" com um aumento característico na expressão superficial de fosfatidilserina (PS) e P-selectina, o que as distingue de plaquetas "apoptóticas" que são positivas apenas para exposição a PS, bem como plaquetas "pró-agregatórias" que são positivas para expressão de P-selectina. Descobrimos que a trombina induz um aumento dose-dependente na proporção de plaquetas pró-coagulantes positivas para a expressão de P-selectina e PS, conforme detectado pela ligação do antic...

Análise abrangente de plaquetas pró-coagulantes exibindo características de necrose, apoptose e ativação plaquetária

Platelets circulating in the bloodstream are relatively quiescent but become "activated" upon encountering stimulants or "agonists" at the site of blood ...

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Medicina

Aqui descrevemos uma técnica experimental de microscopia de fluorescência intravital para visualizar heterotípicos plaquetas neutrófilos interações sobre o endotélio ativado durante a inflamação vascular e formação de trombos em camundongos vivos. Esta tecnologia microscópico será valiosa para estudar o mecanismo molecular da doença vascular e para testar agentes farmacológicos, em condições fisiopatológicas.

Imagens em tempo real de heterotípica plaquetas neutrófilos Interações sobre o endotélio ativado durante a inflamação vascular e formação de trombos em camundongos vivos

Este trabalho descreve um método baseado em microscopia de fluorescência para o estudo da adesão, disseminação e secreção de plaquetas sob fluxo. Esta plataforma versátil permite a investigação da função plaquetária para pesquisas mecanicistas sobre trombose e hemostasia.

Imagens de células vivas de degradação e secreção de plaquetas sob fluxo

A formação de plaquetas pró-coagulantes tem sido correlacionada com um risco aumentado de trombose. Apresenta-se aqui um protocolo preciso para isolar plaquetas lavadas do sangue humano, destinado a quantificar a exposição à fosfatidilserina e a liberação de microvesículas, que são características distintivas das plaquetas pró-coagulantes.

Caracterização de plaquetas pró-coagulantes medindo a exposição à fosfatidilserina e a liberação de microvesículas de plaquetas purificadas humanas

A função plaquetária sob fluxo pode ser avaliada e a ressuscitação hemostática simulada pode ser modelada usando um dispositivo microfluídico, que tem aplicações em trauma e medicina transfusional.

Microfluídica na avaliação da função plaquetária

A citometria de fluxo multicolorida para plaquetas pode identificar novos subtipos de plaquetas dentro de subpopulações clássicas de plaquetas, como plaquetas em repouso, agregatórias, pró-coagulantes e apoptóticas. Isso permite a comparação de diferentes padrões de subpopulação induzidos por vários agonistas plaquetários. Aqui, um procedimento para estabelecer um painel de citometria de fluxo multicolorido é descrito em detalhes.

Análise de subpopulações plaquetárias por citometria de fluxo multicolorida

Este trabalho descreve um ensaio microfluídico que utiliza o estresse de cisalhamento para desencadear a formação de trombos no sangue e caracteriza o trombo por múltiplas dimensões de leitura. O ensaio pode medir a tendência protrombótica de amostras de sangue e, portanto, é útil no diagnóstico de doenças, descoberta de fármacos, bem como em estudos mecanicistas básicos relacionados à trombose.

Ensaio de Perfil de Trombo: Uma Plataforma Baseada em Microfluídica para Caracterização Abrangente da Trombogênese Biomecânica

Um banco de dados de oncologia torácica foi desenvolvido para servir como um repositório completo de dados clínicos e laboratoriais para fins de investigação de translação. O banco de dados servirá pesquisadores de câncer translacional dentro do Programa de Cirurgia Torácica Oncology Research. Este banco de dados é adaptável aos modelos outro câncer, bem como outras doenças humanas.

Geração de banco de dados de Oncologia Torácica Comprehensive - Ferramenta para Pesquisa Translacional

Avaliações objetivas dos mecanismos fisiológicos que suportam discurso são necessárias para controlar o início da doença e progressão em pessoas com esclerose lateral amiotrófica e quantificar os efeitos do tratamento em ensaios clínicos. Neste vídeo, apresentamos uma solução global, protocolo de instrumentação baseada para quantificar o desempenho motor da fala em populações clínicas.

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A técnica de impedância elétrica em tempo real baseado medir Invasão de monocamada de células endoteliais por células cancerosas

RNA de interferência (RNAi) possui muitas vantagens sobre knockout do gene e tem sido amplamente utilizada como uma ferramenta em estudos de genes funcionais. A invenção da tecnologia de RNAi DNA vector baseado fez a longo prazo e knockdown induzível gene possível, e também aumentou a viabilidade de silenciamento do...

DNA Vector baseado em RNA de interferência para estudar a função Gênica em Câncer

Aprenda a analisar plaquetas pró-coagulantes com características de necrose, apoptose e ativação em plaquetas humanas lavadas. Este método auxilia na identificação de biomarcadores para trombose e alvos terapêuticos.

Análise abrangente de plaquetas pró-coagulantes exibindo características de necrose, apoptose e ativação plaquetária

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