Name: Análise por citometria de massa das respostas imunes sistêmicas e locais no carcinoma hepatocelular (Vídeo) | JoVE Journal
Uploaded: 2025-04-25T13:00:00.000+00:00
Duration: 8 min 25 s
Description: Aprenda a usar citometria de massa para analisar respostas imunes sistêmicas e locais no carcinoma hepatocelular. Este método identifica 14 tipos celulares distintos em amostras de sangue periférico, revelando perfis de expressão únicos.

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<table><tbody><tr><td>1&times;PBS</td><td>HyClone</td><td>SH30256.01</td><td></td></tr><tr><td>10&times;PBS</td><td>HyClone</td><td>SH30256.01</td><td></td></tr><tr><td>100 mm&times;20 mm tissue-culture-treated culture dish</td><td>Corning</td><td>430167</td><td></td></tr><tr><td>1000 mL pipette tips</td><td>Rainin</td><td>30389218</td><td></td></tr><tr><td>15 mL centrifuge tube</td><td>NEST</td><td>601052</td><td></td></tr><tr><td>200 mL pipette tips</td><td>Rainin</td><td>30389241</td><td></td></tr><tr><td>40 mm nylon cell strainer/70-mm nylon cell strainer</td><td>Falcon</td><td>352340</td><td></td></tr><tr><td>50 mL centrifuge tube</td><td>NEST</td><td>602052</td><td></td></tr><tr><td>70 &mu;m syringe fifilter</td><td>Sangon Biotech</td><td>F613462-9001</td><td></td></tr><tr><td>Anti-Human CCR2 Antibody (clone: K036C2)</td><td>BioLegend</td><td>357224</td><td></td></tr><tr><td>Anti-Human CCR3 Antibody (clone: 5E8)</td><td>BioLegend</td><td>310724</td><td></td></tr><tr><td>Anti-Human CCR7 Antibody (clone: G043H7)</td><td>BioLegend</td><td>353240</td><td></td></tr><tr><td>Anti-Human CD103 Antibody (clone: Ber-ACT8)</td><td>BioLegend</td><td>350202</td><td></td></tr><tr><td>Anti-Human CD115 Antibody (clone: 9-4D2-1E4)</td><td>BioLegend</td><td>347314</td><td></td></tr><tr><td>Anti-Human CD117 Antibody (clone: 104D2)</td><td>BioLegend</td><td>313201</td><td></td></tr><tr><td>Anti-human CD11b Antibody (clone: 1CRF44)&nbsp;</td><td>BD</td><td>562721</td><td></td></tr><tr><td>Anti-human CD11c Antibody (clone: B-ly6)&nbsp;</td><td>BD</td><td>563026</td><td></td></tr><tr><td>Anti-Human CD123 Antibody (clone: 6H6)</td><td>BioLegend</td><td>306002</td><td></td></tr><tr><td>Anti-Human CD127 Antibody (clone: A019D5)</td><td>BioLegend</td><td>351337</td><td></td></tr><tr><td>Anti-Human CD13 Antibody (clone: WM19)</td><td>BioLegend</td><td>301701</td><td></td></tr><tr><td>Anti-Human CD138 Antibody (clone: MI15)</td><td>BioLegend</td><td>356535</td><td></td></tr><tr><td>Anti-human CD14 Antibody (clone: HCD14)</td><td>BioLegend</td><td>325604</td><td></td></tr><tr><td>Anti-Human CD141 Antibody (clone: M80)</td><td>BioLegend</td><td>344102</td><td></td></tr><tr><td>Anti-Human CD15 Antibody (clone: QA19A61)</td><td>BioLegend</td><td>376302</td><td></td></tr><tr><td>Anti-human CD16 Antibody (clone: B7311)&nbsp;</td><td>BD</td><td>561313</td><td></td></tr><tr><td>Anti-Human CD161 Antibody (clone: HP-3G10)</td><td>BioLegend</td><td>339902</td><td></td></tr><tr><td>Anti-Human CD163 Antibody (clone: GHI/61)</td><td>BioLegend</td><td>333603</td><td></td></tr><tr><td>Anti-Human CD169 Antibody (clone: 7-239)</td><td>BioLegend</td><td>346002</td><td></td></tr><tr><td>Anti-human CD19 Antibody (clone: HIB19)</td><td>BioLegend</td><td>302226</td><td></td></tr><tr><td>Anti-Human CD1c Antibody (clone: L161)</td><td>BioLegend</td><td>331501</td><td></td></tr><tr><td>Anti-Human CD20 Antibody (clone: 2H7)</td><td>BioLegend</td><td>302301</td><td></td></tr><tr><td>Anti-Human CD206 Antibody (clone: 15-2)</td><td>BioLegend</td><td>321151</td><td></td></tr><tr><td>Anti-Human CD24 Antibody (clone: ML5)</td><td>BioLegend</td><td>311129</td><td></td></tr><tr><td>Anti-Human CD25 Antibody (clone: BC96)</td><td>BioLegend</td><td>302624</td><td></td></tr><tr><td>Anti-human CD3 Antibody (clone: UCHT1)&nbsp;</td><td>BD</td><td>555916</td><td></td></tr><tr><td>Anti-Human CD31 Antibody (clone: W18200D)</td><td>BioLegend</td><td>375902</td><td></td></tr><tr><td>Anti-Human CD32 Antibody (clone: FUN-2)</td><td>BioLegend</td><td>303232</td><td></td></tr><tr><td>Anti-Human CD326 Antibody (clone: CO17-1A)</td><td>BioLegend</td><td>369812</td><td></td></tr><tr><td>Anti-Human CD33 Antibody (clone: WM53)</td><td>BioLegend</td><td>303402</td><td></td></tr><tr><td>Anti-human CD4 Antibody (clone: L200)&nbsp;</td><td>BD</td><td>563094</td><td></td></tr><tr><td>Anti-Human CD45 Antibody (clone: HI30)&nbsp;</td><td>BD</td><td>563716</td><td></td></tr><tr><td>Anti-Human CD45RO Antibody (clone: UCHL1)</td><td>BioLegend</td><td>304220</td><td></td></tr><tr><td>Anti-human CD56 Antibody (clone: 5.1H11)</td><td>BioLegend</td><td>362510</td><td></td></tr><tr><td>Anti-Human CD64 Antibody (clone: S18012C)</td><td>BioLegend</td><td>399502</td><td></td></tr><tr><td>Anti-Human CD66b Antibody (clone: 6/40c)</td><td>BioLegend</td><td>392917</td><td></td></tr><tr><td>Anti-human CD68 Antibody (clone: Y1/82A)</td><td>BioLegend</td><td>333808</td><td></td></tr><tr><td>Anti-Human CD69 Antibody (clone: FN50)</td><td>BioLegend</td><td>310902</td><td></td></tr><tr><td>Anti-Human CD7 Antibody (clone: 4H9/CD7)</td><td>BioLegend</td><td>395602</td><td></td></tr><tr><td>Anti-human CD8 Antibody (clone: RPA-T8)&nbsp;</td><td>BD</td><td>557750</td><td></td></tr><tr><td>Anti-Human CD80 Antibody (clone: W17149D)</td><td>BioLegend</td><td>375402</td><td></td></tr><tr><td>Anti-Human CD86 Antibody (clone: BU63)</td><td>BioLegend</td><td>374202</td><td></td></tr><tr><td>Anti-Human FOXP3 Antibody (clone: 206D)</td><td>BioLegend</td><td>320101</td><td></td></tr><tr><td>Anti-Human HLA_ABC Antibody (clone: W6/32)</td><td>BioLegend</td><td>311426</td><td></td></tr><tr><td>Anti-human HLA-DR Antibody (clone: L243)</td><td>BioLegend</td><td>307650</td><td></td></tr><tr><td>Anti-Human IgD Antibody (clone: IA6-2)</td><td>BioLegend</td><td>348211</td><td></td></tr><tr><td>Anti-Human Ki67 Antibody (clone: Ki-67)</td><td>BioLegend</td><td>350501</td><td></td></tr><tr><td>Anti-Human PD_L2 Antibody (clone: MH22B2)</td><td>BD</td><td>567783</td><td></td></tr><tr><td>Anti-Human PD1 Antibody (clone: EH12.2H7)</td><td>BioLegend</td><td>329951</td><td></td></tr><tr><td>Anti-Human PDL1 Antibody (clone: MIH2)</td><td>BioLegend</td><td>393602</td><td></td></tr><tr><td>Anti-human TCR-&gamma;&delta; Antibody (clone: B1)&nbsp;</td><td>BD</td><td>74

análise por citometria de massa das respostas imunes sistêmicas e locais no carcinoma hepatocelular

Mass Cytometry Analysis of Systemic and Local Immune Responses in Hepatocellular Carcinoma

Análise por citometria de massa das respostas imunes sistêmicas e locais no carcinoma hepatocelular

Hepatocellular carcinoma (HCC) is one of the most common and deadliest forms of liver cancer worldwide. Despite advances in treatment, the prognosis for ...

mass-cytometry-analysis-systemic-local-immune-responses

Research

JoVE Journal

Biology

Biologia

Aqui nós descrevemos um método para separar e enriquecer os componentes do microambiente de imune e não imune a tumor em tumores subcutâneos estabelecidas. Esta técnica permite a análise separada de infiltrar imune tumor e frações não imune tumor que podem permitir a caracterização abrangente do microambiente imune do tumor.

Enriquecimento e caracterização do microambiente que imune e não imune Tumor em tumores murino subcutâneos estabelecidas

Aqui nós descrevemos uma abordagem proteômica de célula única para avaliar imunológico fenotípico e funcional (indução de citocinas intracelulares) alterações nas amostras de sangue periférico, analisadas através de citometria de massa.

Análise de célula única do imunofenótipo e produção de citocinas no sangue periférico através de citometria de massa

Aqui, nós descrevemos o desenvolvimento de um modelo murino clìnica relevante do cancro do fígado que recapitulando as características imunes típicas do cancro hepatocelular (HCC).

Um modelo de camundongo de câncer hepatocelular induzido por hepatócitos oncogênico, decorrente do ajuste da inflamação e fibrose hepática

A erradicação mediada por células por células cancerígenas é importante para a iniciação e progressão do câncer. Aqui, apresentamos dois protocolos não baseados em radioatividade para avaliar a citotoxicidade mediada por célulank em relação às células tumorais hepáticas. Além disso, um terceiro protocolo é apresentado para analisar a migração de células NK.

Medição da Citotoxicidade e Migração Mediadas por Células Assassinas Naturais no Contexto das Células Tumorais Hepáticas

Aqui, descrevemos uma estratégia simples e acessível para visualizar, quantificar e mapear células imunes em seções de tecido tumoral parafina fixada em formalina. Essa metodologia combina técnicas de imagem e análise digital existentes com o objetivo de ampliar a capacidade de multiplexação e a análise multiparâmetro de ensaios de imagem.

Visualização, Quantificação e Mapeamento de Populações de Células Imunes no Microambiente tumoral

Modelos in vitro abrangentes que recapitulem fielmente a doença humana relevante estão em falta. O presente estudo apresenta a criação e cultura de spheróides tumorais tridimensionais (3D), uma ferramenta in vitro confiável para estudar a interação tumor-estrommal no carcinoma hepatocelular humano.

Um modelo esferoide tridimensional para investigar a interação tumoral-estroma em carcinoma hepatocelular

Este protocolo descreve a geração de camundongos do sistema imunológico humano (HIS) para estudos de imuno-oncologia. Instruções e considerações sobre o uso deste modelo para testes de imunoterápicos humanos em tumores humanos implantados neste modelo são apresentadas com ênfase na caracterização da resposta do sistema imune humano ao tumor.

Testando imunoterápicos contra o câncer em um modelo humanizado de camundongo com tumores humanos

Aqui, detalhamos um protocolo simples, rápido e confiável assistido por citometria de fluxo multiparamétrico e tumor em um chip para monitorar e caracterizar as etapas que constituem o ciclo de imunidade ao câncer. De fato, uma compreensão completa das interações entre câncer e células imunológicas fornece insights críticos para superar os tumores e orientar o atendimento clínico.

Sistemas de co-cultura in vitro para reproduzir o ciclo de imunidade ao câncer

A presença de mastócitos na margem interna e áreas peritumorais do carcinoma hepatocelular após a ressecção confere um prognóstico favorável. Este estudo endossa o software de análise de imagens QuPath como uma plataforma promissora que pode atender à necessidade de reprodutibilidade, consistência e precisão em patologia digital.

Mastócitos no microambiente do carcinoma hepatocelular conferem prognóstico favorável: um estudo retrospectivo usando o software de análise de imagem QuPath 

Este vídeo demonstra a dissecção e remoção do timo fetal, bem como o preparo de culturas ex vivo de 2-dGuo tratados com timo.

Preparação de 2-dGuo-Tratada Culturas Organ Thymus

Aprenda a usar citometria de massa para analisar respostas imunes sistêmicas e locais no carcinoma hepatocelular. Este método identifica 14 tipos celulares distintos em amostras de sangue periférico, revelando perfis de expressão únicos.

Análise por citometria de massa das respostas imunes sistêmicas e locais no carcinoma hepatocelular

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