Name: Análisis exhaustivo de plaquetas procoagulantes que presentan características de necrosis, apoptosis y activación plaquetaria (Video) | JoVE Journal
Uploaded: 2025-05-23T14:00:00.000+00:00
Duration: 4 min 37 s
Description: Aprende a analizar plaquetas procoagulantes con características de necrosis y apoptosis usando citometría de flujo. Este método permite identificar biomarcadores y dianas terapéuticas para estados protrombóticos.

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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>Prodan, C. I., Vincent, A. S., Dale, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coated+platelet+levels+correlate+with+bleed+volume+in+patients+with+spontaneous+intracerebral+hemorrhage%5BTitle%5D)+AND+%22Stroke%22%5BJournal%5D)">Coated platelet levels correlate with bleed volume in patients with spontaneous intracerebral hemorrhage.</a> Stroke. 41 (6), 1301-1303 (2010).</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>
<li>Choo, H. -J., Saafir, T. B., Mkumba, L., Wagner, M. B., Jobe, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+calcium+and+reactive+oxygen+species+regulate+agonist-initiated+platelet+phosphatidylserine+exposure%5BTitle%5D)+AND+%22Arterioscler+Thromb+Vasc+Biol%22%5BJournal%5D)">Mitochondrial calcium and reactive oxygen species regulate agonist-initiated platelet phosphatidylserine exposure.</a> Arterioscler Thromb Vasc Biol. 32 (12), 2946-2955 (2012).</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>
<li>Goelz, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Platelets+express+adaptor+proteins+of+the+extrinsic+apoptosis+pathway+and+can+activate+caspase-8%5BTitle%5D)+AND+%22PLOS+ONE%22%5BJournal%5D)">Platelets express adaptor proteins of the extrinsic apoptosis pathway and can activate caspase-8.</a> PLOS ONE. 16 (1), e0244848(2021).</li>
<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>
<li>Nadine, J. A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coated+platelets+function+in+platelet-dependent+fibrin+formation+via+integrin+%CE%B1IIb%CE%B23+and+transglutaminase+factor+XIII%5BTitle%5D)+AND+%22Haematologica%22%5BJournal%5D)">Coated platelets function in platelet-dependent fibrin formation via integrin αIIbβ3 and transglutaminase factor XIII.</a> Haematologica. 101 (4), 427-436 (2016).</li>
<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>Solución de citrato ácido dextrosa (ACD) (para 1000 mL)</td><td></td><td></td><td>Citrato de tri-sodio- &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 Dextrosa-&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 Agua-&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; Volumen de maquillaje hasta 1000 mL&nbsp;</td></tr><tr><td>Alexa Fluor 488 kit de etiquetado de proteínas</td><td>Invitrogen</td><td>A10235</td><td></td></tr><tr><td>APC Ratón Anti-Humano 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>Tampón A (Preparación 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. en 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 100 mL de solución 10X HEPES&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238.30 y 230 &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 y 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 Agua a 100 mL después de ajustar el pH a 6,2 Diluya la solución 10X 1:10 (v/v) con agua Milli Q justo antes de la preparación de las plaquetas para obtener la solución 1X que debe complementarse con lo siguiente &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. en 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 1 ml de solución 1X EGTA &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Suma 10 &mu; L (1:100 v/v) 100 mM EGTA&nbsp; Glucosa &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 mM &nbsp; &nbsp; &nbsp; &nbsp; Suma 5 &mu; L (1:200 v/v) 1 M glucosa PGE1 &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3 y mu; M &nbsp; &nbsp; &nbsp; &nbsp; Suma 3 &mu; L (1:333 v/v) 1 mM solución PGE1</td></tr><tr><td>Tampón B (Preparación 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. en 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 100 mL de solución 10X HEPES&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238.30 y 230 &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 y 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 Agua a 100 mL después de ajustar el pH a 7,4 Diluya la solución 10X 1:10 (v/v) con agua Milli Q justo antes de la preparación de las plaquetas para obtener la solución 1X que debe complementarse con lo siguiente &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. en 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Para 1 ml 1X solución Glucosa &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 mM &nbsp; &nbsp; &nbsp; &nbsp; Suma 5 &mu; L (1:200 v/v) 1 M de glucosa&nbsp;</td></tr><tr><td>CaCl2 (0,5 M) (Para 50 ml)</td><td></td><td></td><td>Peso molecular de CaCl2.2H2O = 147,02 Disolver 3,675 g de CaCl2,2H2O en 50 ml de agua Milli Q</td></tr><tr><td>Reactivos de detección CellEvent Caspasa-3/7 Green</td><td>Invitrogen</td><td>C10423</td><td>Reactivo de detección de apoptosis</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 de EDTA Na2.2H2 O: 372.24g Pesar 18.612 g y suspender en 50 ml de agua Milli Q y verificar el pH (pH~4) Agregue lentamente 10N NaOH agitando y controle el pH. EDTA comienza a solubilizar alrededor de pH 7 y es completamente soluble a pH 8. Aumente el volumen hasta 100 ml con agua 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 Agregue 3.804 g de EGTA en 50 ml de agua Milli Q y verifique el pH (pH~3) Agregue 10 N de NaOH gota a gota mientras agita y controle el pH EGTA se vuelve soluble a pH 7.0 (aprox) Ajuste el pH a 7.4 Aumente el volumen a 100 ml con agua</td></tr> <tr><td>Milli QFITC Ratón Anti-Humano PAC-1</td><td>BD 340507</td><td></td><td></td></tr><tr><td>FITC-IETD-FMK Caspasa 8 (activo) kit de tinción</td><td>Abcam</td><td>ab65614</td><td></td></tr><tr><td>Mitotracker Red CMXRos (colorante de marcado de mitochindrias)</td><td>Invitrogen</td><td>M7512</td><td>Stock= 1 mM (50 &micro; g disuelto en 90 &micro; l DMSO) Subexistencias= 100 &micro; M (10 y micro; l Stock + 90 y micro; l DMSO) Concentración de trabajo = 500nM (0.5 &micro; l en 100 y 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>Stock= 20 mM (1 mg disuelto en 141 &micro; L DMSO) Substock= 1 mM (10 &micro; l Stock + 190 y micro; L DMSO) Concentración de trabajo= 3 &micro; M (3 y micro; L en 1 mL)</td></tr><tr><td>Rhod-2 AM</td><td>Invitrogen</td><td>R1244</td><td>Stock= 5 mM (1 mg disuelto en 178 &micro; L DMSO) Subexistencias= 100 &micro; M (10 y micro; L Stock + 90 &micro; L DMSO) Concentración de trabajo= 500 nM (0,5 &micro; L en 100 y micro; L)</td></tr> <tr><td>Trombina de plasma humano</td><td>Sigma</td><td>T7572</td><td></td></tr></tbody></table>

análisis exhaustivo de plaquetas procoagulantes que presentan características de necrosis, apoptosis y activación plaquetaria

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

Una proporción de plaquetas activadas se vuelven "procoagulantes" con un aumento característico en la expresión superficial tanto de fosfatidilserina (PS) como de P-selectina, lo que las distingue de las plaquetas "apoptóticas" que son positivas solo para la exposición a PS, así como de las plaquetas "proagregadoras" que son positivas para la expresión de P-selectina. Encontramos que la trombina induce un aumento dependiente de la dosis en la proporción de plaquetas procoagulantes positivas tanto para la expresión de P-s...

Análisis exhaustivo de plaquetas procoagulantes que presentan características de necrosis, apoptosis y activación plaquetaria

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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Aquí se presenta una técnica experimental de microscopía intravital de fluorescencia para visualizar heterotípicas plaquetas neutrófilos interacciones en el endotelio activado durante la inflamación vascular y la formación de trombos en ratones vivos. Esta tecnología microscópica será valiosa para estudiar el mecanismo molecular de la enfermedad vascular y para poner a prueba los agentes farmacológicos bajo condiciones fisiopatológicas.

Imágenes en tiempo real de heterotypic plaquetas neutrófilos Interacciones en el endotelio activado durante la inflamación vascular y la formación de trombos en ratones vivos

Este trabajo describe un método basado en microscopía de fluorescencia para el estudio de la adhesión plaquetaria, la extensión y la secreción bajo flujo. Esta plataforma versátil permite la investigación de la función plaquetaria para la investigación mecánica sobre trombosis y hemostasia.

Imágenes de células vivas de desgranulación plaquetaria y secreción bajo flujo

La formación de plaquetas procoagulantes se ha correlacionado con un mayor riesgo de trombosis. Aquí se presenta un protocolo preciso para aislar plaquetas lavadas de la sangre humana, destinado a cuantificar la exposición a la fosfatidilserina y la liberación de microvesículas, que son características distintivas de las plaquetas procoagulantes.

Caracterización de plaquetas procoagulantes mediante la medición de la exposición a fosfatidilserina y la liberación de microvesículas de plaquetas humanas purificadas

Se puede evaluar la función plaquetaria bajo flujo y se puede modelar la reanimación hemostática simulada utilizando un dispositivo microfluídico, que tiene aplicaciones en medicina traumatológica y transfusional.

Microfluídica en la evaluación de la función plaquetaria

La citometría de flujo multicolor para plaquetas puede identificar nuevos subtipos de plaquetas dentro de las subpoblaciones de plaquetas clásicas, como las plaquetas en reposo, agregativas, procoagulantes y apoptóticas. Esto permite la comparación de diferentes patrones de subpoblación inducidos por varios agonistas plaquetarios. Aquí, se describe en detalle un procedimiento para establecer un panel de citometría de flujo multicolor.

Análisis de subpoblaciones de plaquetas mediante citometría de flujo multicolor

Este trabajo describe un ensayo microfluídico que utiliza el esfuerzo cortante para desencadenar la formación de trombos en sangre y caracteriza el trombo mediante múltiples dimensiones de lectura. El ensayo puede medir la tendencia protrombótica de las muestras de sangre y, por tanto, es útil en el diagnóstico de enfermedades, el descubrimiento de fármacos, así como en estudios mecanicistas básicos relacionados con la trombosis.

Ensayo de Perfilado de Trombo: Una plataforma basada en microfluídica para caracterizar de forma exhaustiva la trombogénesis biomecánica

Una base de datos de oncología torácica fue desarrollado para servir como un repositorio global de datos clínicos y de laboratorio para los fines de la investigación traslacional. La base de datos servirá investigadores traslacional del cáncer dentro del Programa de Oncología Torácica de Investigación. Esta base de datos se adapta a los modelos de otros tipos de cáncer, así como otras enfermedades humanas.

Generación de base de datos integral de Oncología Torácica - Herramienta para la Investigación Traslacional

Evaluaciones objetivas de los mecanismos fisiológicos que el apoyo del habla son necesarias para controlar la enfermedad de inicio y la progresión de las personas con ELA y para cuantificar los efectos del tratamiento en los ensayos clínicos. En este vídeo, se presenta una completa instrumentación basada en el protocolo para cuantificar el rendimiento motor del habla en poblaciones clínicas.

Un Protocolo de Evaluación Integral de la disfunción bulbar de la Esclerosis Lateral Amiotrófica (ALS)

A tiempo real Técnica de Impedancia Eléctrica base para medir la invasión de monocapa de células endoteliales de las células cancerosas

ARN de interferencia (RNAi) posee muchas ventajas sobre los octavos de final de genes y ha sido ampliamente utilizado como una herramienta en los estudios de genes funcionales. La invención de ADN basado en vectores RNAi la tecnología ha hecho a largo plazo y caída de genes inducibles posible, y también aumentó la viabilidad de silenciamiento de los...

ADN vectorial basado en el ARN de interferencia para estudiar la función génica en el cáncer

Aprende a analizar plaquetas procoagulantes con características de necrosis y apoptosis usando citometría de flujo. Este método permite identificar biomarcadores y dianas terapéuticas para estados protrombóticos.

Análisis exhaustivo de plaquetas procoagulantes que presentan características de necrosis, apoptosis y activación plaquetaria

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