Name: Isolamento de Núcleo Único de Tecido de Endarterectomia Coronária de Pacientes com Bypass de Artéria Coronária (Vídeo) | JoVE Journal
Uploaded: 2026-04-03T13:08:00.000+00:00
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Description: Aprenda a isolar núcleos únicos de tecido de endarterectomia coronária de pacientes com bypass de artéria coronária. Este método preserva a integridade nuclear para sequenciamento de RNA e análise transcriptômica de placas ateroscleróticas instáveis.

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<table><tbody><tr><td>100 & mu; Filtro m</td><td>Miltenyi Biotech&nbsp;</td><td>130-098-463</td><td></td></tr><tr><td>30 & mu; Filtro m</td><td>Miltenyi Biotech&nbsp;</td><td>130-098-458</td><td></td></tr><tr><td>Albumina Sérica Bovina</td><td>HiMedia</td><td>TC546-25G</td><td></td></tr><tr><td>Centrífuga</td><td>MPW</td><td>NA</td><td>https://mpw.pl/en/offer/mpw-352r</td></tr><tr><td>Microscópio Composto</td><td>Nikon</td><td>NA&nbsp;</td><td>Microscópio invertido Nikon Eclipse Ts2 (https://www.microscope.healthcare.nikon.com/en_AOM/products/inverted-microscopes/eclipse-ts2)</td></tr><tr><td>Microscópio Confocal &nbsp;</td><td>Nikon</td><td>NA</td><td>Microscópio confocal invertido Nikon Ti2-E (https://www.microscope.healthcare.nikon.com/en_AOM/products/inverted-microscopes/eclipse-ti2-series)</td></tr><tr><td>Coberturas</td><td>HiMedia</td><td>BG011C</td><td></td></tr><tr><td>Frasco de Amostra CRYOCHIL de Boca Larga</td><td>Tarsons</td><td>883191</td><td></td></tr><tr><td>DAPI</td><td>Sigma</td><td>D9542-5MG</td><td></td></tr><tr><td>DPBS</td><td>Gibco (Termofischer)</td><td>10010-023</td><td></td></tr><tr><td>Tubo de centrífuga (15 mL)</td><td>Tarsons</td><td>546021</td><td></td></tr><tr><td>Citômetro de fluxo &nbsp;</td><td>BD em Biociências</td><td>663029</td><td>BD FACSLyric (3L12C Instrumento CEIVD)</td></tr><tr><td>Hemocitômetro</td><td>Axibio</td><td>CCB 100</td><td></td></tr><tr><td>Tubo de Microcentrífuga (1,5 mL)</td><td>Tarsons</td><td>500010C</td><td></td></tr><tr><td>Tubo Microcentrífuga (5 mL)</td><td>Tarsons</td><td>500050</td><td></td></tr><tr><td>MgCl2</td><td>Sigma</td><td>M1028-100ML</td><td></td></tr><tr><td>Micro pilão</td><td>Tarsons</td><td>160020</td><td></td></tr><tr><td>NaCl</td><td>Sigma</td><td>S9888-500G</td><td></td></tr><tr><td>NP-40</td><td>HiMedia</td><td>MB143-100ML</td><td></td></tr><tr><td>Água livre de nuclease</td><td>Gibco (Termofischer)</td><td>W4502-1L</td><td></td></tr><tr><td>Inibidor de RNase</td><td>Sigma</td><td>333540201</td><td></td></tr><tr><td>RPMI 1640</td><td>Gibco (Termofischer)</td><td>11875093</td><td></td></tr><tr><td>Slide&nbsp;</td><td>Riviera</td><td>72910135</td><td></td></tr><tr><td>Tris-HCl</td><td>Sigma</td><td>T2194-100ML</td><td></td></tr><tr><td>Azul de tripan & nbsp;</td><td>Sigma</td><td>302643-25G</td><td></td></tr></tbody></table>

isolamento de núcleo Único de tecido de endarterectomia coronária de pacientes com bypass de artéria coronária

Single Nuclei Isolation from Coronary Endarterectomy Tissue of Coronary Artery Bypass Graft Patients

O tecido de endarterectomia coronária humana foi coletado de pacientes submetidos à cirurgia de CABG. O tecido foi pesado e processado para isolamento de núcleos, e núcleos mais isolados foram confirmados por citometria de fluxo e microscopia confocal (Figura 1).
Avaliação do azul de tripano da qualidade do isolamento dos núcleos O tecido mantido em médio RPMI foi pesado e mediu 247,8 mg. O tecido foi triturado e depois homogeneizado em tampão ...

Isolamento de Núcleo Único de Tecido de Endarterectomia Coronária de Pacientes com Bypass de Artéria Coronária

Atherosclerosis is a chronic, progressive inflammatory condition characterized by the accumulation of plaque in the walls of arteries. During the ...

single-nuclei-isolation-from-coronary-endarterectomy-tissue-coronary

Research

JoVE Journal

Biology

Biologia

Núcleos cardíaca são isolados através de sedimentação densidade e immunolabeled com anticorpos contra o material pericentriolar 1 (PCM-1) para identificar e classificar os núcleos dos cardiomiócitos por citometria de fluxo.

Isolamento dos núcleos dos cardiomiócitos a partir de Post-mortem de tecidos

O conhecimento atual sobre a base celular de doenças cardíacas se baseia principalmente em estudos em modelos animais. Aqui nós descrevemos e validar um novo método para obter simples cardiomiócitos viáveis ​​a partir de pequenas amostras cirúrgicas do miocárdio ventricular humano. Miócitos ventriculares humanos podem ser usados ​​para estudos eletrofisiológicos e testes de drogas.

Isolamento e caracterização funcional de Ventricular Humano cardiomiócitos a partir de amostras cirúrgicas frescos

A capacidade de isolar células endoteliais da válvula cardíaca (VECs) é fundamental para a compreensão dos mecanismos de desenvolvimento, manutenção e doença da válvula.  Aqui descrevemos o isolamento de VECs de camundongos Tie2-GFP embrionários e adultos usando FACS que permitirá estudos determinando a contribuição de VECs em processos de desenvolvimento e doença.

O isolamento de células endoteliais de Murino Válvula

O objectivo deste protocolo é demonstrar as técnicas de isolamento e cultivo de células musculares murino primários liso vascular (VSMC) da circulação coronária. Uma vez que as VSMCs foram isolados, eles podem ser usados ​​para várias técnicas de cultura convencionais.

Isolamento de Murino vasculares coronários células musculares lisas

Este protocolo está preparado para compartilhar nosso método de isolamento de células endoteliais coronárias de camundongos para fins de imagem ou para conduzir experimentos biológicos moleculares.

Isolamento de células de camundongo Coronária endoteliais

tecido cardíaco humano abriga populações de células precursoras perivasculares multipotentes que podem ser adequados para a regeneração do miocárdio. A técnica aqui descrita permite o isolamento e purificação simultânea de duas populações de células estromais multipotentes associados com os vasos nativos de sangue, ie CD146 + CD34 - pericitos e células CD34 + CD146 - células adventícias, a partir do miocárdio humano.

Isolamento de perivascular Multipotentes populações de células precursoras a partir de tecido cardíaco humano

Células endoteliais de tumor são importantes determinantes do microambiente do tumor e o curso da doença. Aqui, um protocolo para o isolamento de células endoteliais puras e viáveis de carcinoma colorectal humana e cólon normal para ser usado em pesquisa de teste e patogênese da droga é descrito.

Isolamento de células endoteliais humanas do cólon Normal e Carcinoma colorrectal - um protocolo melhorado

O objetivo deste trabalho é desenvolver um método para isolar os cardiomiócitos do coração adulto e medir o conteúdo e a nucleação do DNA.

Isolamento de Cardiomiócitos de Corações Fixos para Imunocitoquímica e Análise ploidy

Apresentamos um protocolo para o isolamento de células endocárdicas e coronárias de corações de ratos através da digestão sequencial de tecidos em um tampão de digestão, coleta celular de ciclos de centrífuga recorrentes e purificação celular usando microesferas CD31 anti-ratos.

Isolamento das Células Endocárdicas e Coronárias da Parede Livre Ventricular do Coração de Rato

Aqui, apresentamos um protocolo descrevendo a preparação de núcleos celulares. Após microdissecção e dissociação enzimática do tecido cardíaco em células únicas, as células progenitoras foram congeladas, seguidas pelo isolamento de células viáveis puras, que foram usadas para sequenciamento de RNA de núcleo único e o ensaio de núcleo único para cromatina acessível à transposase com análises de sequenciamento de alto rendimento.

Isolamento de Núcleos de Células Progenitoras Cardíacas de Camundongos para Perfil de Epigenoma e Expressão Gênica em Resolução Unicelular

Este estudo apresenta um modelo cirúrgico porcino de isquemia miocárdica crônica por estenose progressiva da artéria coronária, resultando em comprometimento da função cardíaca sem infarto. Após a isquemia, os animais são submetidos à cirurgia de revascularização do miocárdio sem circulação extracorpórea com colocação epicárdica de adesivo carregado de exossomos derivado de células-tronco. Essa terapia adjuvante melhora a função e a recuperação miocárdica.

Modelo cirúrgico porcino de isquemia miocárdica crônica tratada com adesivo carregado de exossomo e cirurgia de revascularização miocárdica sem circulação extracorpórea

Aprenda a isolar núcleos únicos de tecido de endarterectomia coronária de pacientes com bypass de artéria coronária. Este método preserva a integridade nuclear para sequenciamento de RNA e análise transcriptômica de placas ateroscleróticas instáveis.

Isolamento de Núcleo Único de Tecido de Endarterectomia Coronária de Pacientes com Bypass de Artéria Coronária

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