Name: Enxertos adiposos brancos e marrons: uma abordagem para corrigir déficits reprodutivos, metabólicos e renais em camundongos obesos com braquiúria preta e castanha (BTBR) (Vídeo) | JoVE Journal
Uploaded: 2025-09-09T13:45:00.000+00:00
Duration: 6 min 16 s
Description: Aprenda o protocolo padronizado de transplante combinado de tecido adiposo branco e marrom em camundongos BTBR obesos deficientes em leptina. Este método restaura a fertilidade, melhora a função metabólica e renal, oferecendo uma abordagem reprodutível para estudar doenças cardiorrenais metabólicas.

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5 minutes

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</ol>

<table><tbody><tr><td>Solução alcoólica de clorexidina digluconato a 0,5%</td><td>Rioqu&iacute; Mica</td><td>SKU 1068253</td><td></td></tr><tr><td>Agulha calibre 16</td><td>Becton Dickinson</td><td>SKU 305198</td><td></td></tr><tr><td>Solução de digluconato de clorexidina não alcoólica a 2,0%</td><td>Rioqu&iacute; Mica</td><td>SKU 1113045</td><td></td></tr><tr><td>Seringa de 3 mL</td><td>Becton Dickinson</td><td>SKU 309657</td><td></td></tr><tr><td>Fitas Accu-Check e ndash; Performa</td><td>Roche</td><td>EAN 4015630981960</td><td></td></tr><tr><td>Alcian Blue PAS Kit</td><td>EasyPath</td><td>SKU EP-11-20019</td><td></td></tr><tr><td>Albumina Sérica Bovina (BSA)</td><td>Sigma-Aldrich</td><td>A4612</td><td></td></tr><tr><td>Software CellSens</td><td>Olympus</td><td>CELLSENS</td><td></td></tr><tr><td>Citrato Buffer pH 6,0: Ácido cítrico monohidratado + Hidróxido de sódio</td><td>Sigma-Aldrich</td><td>C1909 + 28-3010</td><td></td></tr><tr><td>Anticorpo Caspase-3 Cortado</td><td>Tecnologia de Sinalização Celular</td><td>9661</td><td></td></tr><tr><td>Analisador Cobas C 111</td><td>Roche</td><td>4528778001</td><td></td></tr><tr><td>Creatinina K</td><td>Teste de laboratório</td><td>96-300</td><td></td></tr><tr><td>EnVision FLEX DAB+ Cromogênio</td><td>Agilent-Dako</td><td>K3468</td><td></td></tr><tr><td>Reagente de detecção EnVision FLEX / HRP</td><td>Agilent-Dako</td><td>SM802</td><td></td></tr><tr><td>Reagente Bloqueador de Peroxidase Envision FLEX</td><td>Agilent-Dako</td><td>SM801</td><td></td></tr><tr><td>EnVision FLEX Substrate Buffer</td><td>Agilent-Dako</td><td>SM803</td><td></td></tr><tr><td>O EnVision FLEX foi buffer 20x</td><td>Agilent-Dako</td><td>K800721-2</td><td></td></tr><tr><td>Eosina</td><td>Agilent-Dako</td><td>CS701</td><td></td></tr><tr><td>Leitor de Microplacas GloMax Discover</td><td>Promega</td><td>GM3000</td><td></td></tr><tr><td>Glicosímetro Accu-Check - Performa</td><td>Roche</td><td>EAN 4015630980512</td><td></td></tr><tr><td>Software GraphPad Prism 8 (software estatístico)</td><td>GraphPad por Dotmatics</td><td>-</td><td></td></tr><tr><td>100% v/v Isoforina (Isoflurane)</td><td>Crist&aacute; Lia</td><td>-</td><td></td></tr><tr><td>Hematoxilina</td><td>Agilent-Dako</td><td>CS700</td><td></td></tr><tr><td>Microscópio óptico IX51</td><td>Olympus</td><td>IX51</td><td></td></tr><tr><td>Gaiola Metabólica para camundongos individuais</td><td>Tecniplast</td><td>3600M021</td><td></td></tr><tr><td>Meio de montagem - Entellan</td><td>Sigma-Aldrich</td><td>100869</td><td></td></tr><tr><td>Mouse Albumin ELISA Kit</td><td>Abcam</td><td>Ab207620</td><td></td></tr><tr><td>Cepa do camundongo: BTBR. Cg-Lepob/WiscJ</td><td>Laboratórios JAX</td><td>4824</td><td></td></tr><tr><td>Kit de Schiff Ácido Periódico (PAS)</td><td>EasyPath</td><td>EP-11-20014</td><td></td></tr><tr><td>RM255 Microtome</td><td>Leica Biosystems</td><td>14050237960</td><td></td></tr><tr><td>Processador de tecido TP1020</td><td>Leica Biosystems</td><td>TP-1020</td><td></td></tr><tr><td>Tramadol 50 mg/mL</td><td>Gr&uuml; Nenthal</td><td>4022444</td><td></td></tr><tr><td>Anticorpo WT-1</td><td>Santa Cruz</td><td>SC-192</td><td></td></tr></tbody></table>

enxertos adiposos brancos e marrons: uma abordagem para corrigir déficits reprodutivos, metabólicos e renais em camundongos obesos com braquiúria preta e castanha (btbr)

White and Brown Adipose Grafts: An Approach to Correct Reproductive, Metabolic, and Renal Deficits in Black and Tan Brachyury (BTBR) Obese Mice

Enxertos adiposos brancos e marrons: uma abordagem para corrigir déficits reprodutivos, metabólicos e renais em camundongos obesos com braquiúria preta e castanha (BTBR)

Transplantation of white adipose tissue (WAT) and brown adipose tissue (BAT) has emerged as a promising therapeutic strategy for reversing metabolic ...

white-brown-adipose-grafts-an-approach-to-correct-reproductive

Research

JoVE Journal

Biology

Biologia

Neste vídeo, demonstramos o procedimento cirúrgico para isolar o tecido adiposo branco subcutâneo e o tecido adiposo marrom interescapular de um camundongo recém-nascido.

Coleção de depósito adiposo branco e marrom do filhote de rato: um procedimento cirúrgico para colher o tecido adiposo branco e o tecido adiposo marrom do filhote de camundongo

Principais pré-adipócitos brancos isolados de tecidos adiposos brancos em ratos podem ser diferenciadas em células bege / brite. Apresentado aqui é um sistema modelo celular fiável para estudar a regulação molecular de "escurecimento" da gordura branca.

Isolamento e diferenciação de células estromais vasculares para Células Bege / Brite

Este artigo descreve um método de transplante para enxertar células epiteliais mamárias doadoras na almofada de gordura branca interscapular de animais receptores. Este método pode ser usado para examinar os efeitos do hospedeiro e/ou doador no desenvolvimento do epitélio mamário e elimina a necessidade de pré-compensação, estendendo assim a utilidade desta técnica.

Transplante de células epiteliais mamárias no Bloco de Gordura Branca Interscapular

Este relatório descreve um protocolo para o isolamento simultâneo de preadipócitos primários marrons e brancos de camundongos recém-nascidos. Células isoladas podem ser cultivadas na cultura e induzidas a se diferenciar em adipócitos brancos e marrons totalmente maduros. O método permite a caracterização genética, molecular e funcional das células de gordura primárias na cultura.

Isolamento e diferenciação de preadipócitos brancos e marrons primários de camundongos recém-nascidos

Imagem funcional e quantitação de depósitos de adiposos termogênicos em camundongos usando uma abordagem baseada em imagens micro-PET/MR.

Visualização e Quantificação de Tecidos Adiposos Marrons e Bege em Camundongos usando [18F]FDG Micro-PET/MR Imaging

Os pré-adipócitos são isolados da fração vascular estromal do tecido adiposo marrom interescapular de camundongos recém-nascidos e diferenciados em células que acumulam gotículas lipídicas, expressam marcadores moleculares e mostram a morfologia mitocondrial de adipócitos marrons maduros. Essas células são posteriormente analisadas por imunofluorescência e microscopia eletrônica de transmissão.

Diferenciação e imagem de adipócitos marrons da fração vascular estromal do tecido adiposo interescapular de camundongos recém-nascidos

Aqui, apresentamos um protocolo detalhado delineando o uso de fragmentos microvasculares isolados de tecido adiposo humano ou de roedores como uma abordagem direta para a engenharia de tecido adiposo bege vascularizado funcional.

Cultura Tridimensional de Tecido Adiposo Termogênico Vascularizado a partir de Fragmentos Microvasculares

Este protocolo apresenta o uso do CRISPR SunTag-p65-HSF1 (SPH) em adipócitos (AdipoSPH) como uma estratégia alternativa ao vírus adenoassociado (AAV) para investigar a biologia da gordura bege. A injeção in vivo de sgRNA carreador de AAV visando o gene endógeno Prdm16 é suficiente para induzir o desenvolvimento de gordura bege e melhorar o programa do gene termogênico.

Investigação da Biologia e Metabolismo da Gordura Bege Utilizando o Sistema de Ativação CRISPR SunTag-p65-HSF1

Neste protocolo, descrevemos os procedimentos técnicos para gerar camundongos knockout específicos para tecido adiposo marrom (BAT) utilizando um sistema combinado de RNA guia único (sgRNA) Cre-LoxP, CRISPR-Cas9 e vírus adenoassociado (AAV). As etapas descritas incluem o planejamento dos sgRNAs, a preparação das partículas de AAV-sgRNA e a microinjeção de AAV nos lóbulos BAT.

Geração de camundongos knockout específicos para gordura marrom usando um sistema combinado de RNA guia único Cre-LoxP, CRISPR-Cas9 e vírus adenoassociado

Genética induzível Destino Mapping (GIFM) marcas e faixas de células com o controle espacial e temporal...

Uma Abordagem Prática para mapeamento genético induzível Fate: Um Guia Visual para Mark e Track Células In Vivo</em

Aprenda o protocolo padronizado de transplante combinado de tecido adiposo branco e marrom em camundongos BTBR obesos deficientes em leptina. Este método restaura a fertilidade, melhora a função metabólica e renal, oferecendo uma abordagem reprodutível para estudar doenças cardiorrenais metabólicas.

Enxertos adiposos brancos e marrons: uma abordagem para corrigir déficits reprodutivos, metabólicos e renais em camundongos obesos com braquiúria preta e castanha (BTBR) (Vídeo) | JoVE Journal

Enxertos adiposos brancos e marrons: uma abordagem para corrigir déficits reprodutivos, metabólicos e renais em camundongos obesos com braquiúria preta e castanha (BTBR)

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