Name: Estabelecimento de Culturas Primárias Enriquecidas em Zona a partir do córtex adrenal do camundongo (Vídeo) | JoVE Journal
Uploaded: 2026-05-08T14:20:00.000+00:00
Duration: 7 min 43 s
Description: Aprenda a estabelecer culturas primárias enriquecidas em zona a partir do córtex adrenal do camundongo. Este método permite isolar células da zona glomerulosa e zona fasciculata para estudos de fisiologia adrenal.

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</ol>

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Plate</td><td>Corning (Corning, NY, USA)</td><td>3527</td><td>Tratamento para cultura de tecidos</td></tr><tr><td>Sterile Filter Paper</td><td>N/A</td><td>N/A</td><td>Remoção de gordura e suporte para dissecação</td></tr><tr><td>DMEM/F-12 Medium</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>11320033</td><td>Formulação 1:1</td></tr><tr><td>Fetal Bovine Serum (FBS)</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>10082147</td><td>Inativado por calor</td></tr><tr><td>ITS Supplement</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>41400045</td><td>100X</td></tr><tr><td>GlutaMAX Supplement</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>35050061</td><td>100X</td></tr><tr><td>Penicillin-Streptomycin (100X)</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>15140122</td><td>Antibiótico</td></tr><tr><td>PBS 1X</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>10010023</td><td>pH 7,4</td></tr><tr><td>Collagenase Type I</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>17100-017</td><td>Digestão de tecidos</td></tr><tr><td>DNase I</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>DN25-1</td><td>Digestão de tecidos</td></tr><tr><td>Angiotensin II (Ang II)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>A9525</td><td>Estimulação hormonal</td></tr><tr><td>Adrenocorticotropic Hormone (ACTH)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>A0298</td><td>Estimulação hormonal</td></tr><tr><td>Forskolin</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>F6886</td><td>Estimulação de cAMP</td></tr><tr><td>Potassium Chloride (KCl)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>P9541</td><td>Estimulação de aldosterona</td></tr><tr><td>Sodium Chloride (NaCl)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>S7653</td><td>Controle do veículo</td></tr><tr><td>Small Surgical Scissors</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>14060-09</td><td>Dissecação</td></tr><tr><td>Scalpel with Disposable Blade</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>10003-12</td><td>Microdissecação</td></tr><tr><td>Fine Forceps</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>11252-00</td><td>Dissecação</td></tr><tr><td>Curved Forceps</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>11251-10</td><td>Dissecação</td></tr><tr><td>Laminar Flow Hood</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Processamento estéril de tecidos</td></tr><tr><td>Incubator</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Cultivo de células</td></tr><tr><td>Centrifuge</td><td>Eppendorf (Hamburg, Germany)</td><td>N/A</td><td>Pelletagem de células</td></tr><tr><td>Neubauer Counting Chamber</td><td>Hausser Scientific (Horsham, PA, USA)</td><td>1490</td><td>Contagem de células</td></tr><tr><td>RIPA Lysis and Extraction Buffer</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>89901</td><td>Extração de proteínas</td></tr><tr><td>Halt Protease Inhibitor Cocktail</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>87786</td><td>Extração de proteínas</td></tr><tr><td>Fisherbrand Model 120 Sonic Dismembrator</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>FB120</td><td>Sonicação</td></tr><tr><td>RIA System</td><td>Tecan (M&auml;nnedorf, Switzerland)</td><td>MG13051</td><td>Quantificação hormonal</td></tr><tr><td>Cobra II Auto-Gamma Counter</td><td>Berthold Technologies (Bad Wildbad, Germany)</td><td>N/A</td><td>Medição de radioatividade</td></tr><tr><td>Corticosterone ELISA Kit (96-well)</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>55-CORMS-E01</td><td>Medição hormonal</td></tr><tr><td>ProLong Gold Antifade Mountant</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>P36930</td><td>Meio de montagem</td></tr><tr><td>Mouse anti-Dab2</td><td>BD Biosciences (San Jose, CA, USA)</td><td>610464</td><td>Anticorpo primário</td></tr><tr><td>Chicken anti-GFP</td><td>Aves Labs (Tigard, OR, USA)</td><td>GFP-1020</td><td>Anticorpo primário</td></tr><tr><td>Rabbit anti-RFP</td><td>Rockland Immunochemicals (Limerick, PA, USA)</td><td>600-401-379</td><td>Anticorpo primário</td></tr><tr><td>Alexa Fluor 594 Goat anti-Mouse</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-11005</td><td>Anticorpo secundário</td></tr><tr><td>Alexa Fluor 594 Goat anti-Chicken</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-11042</td><td>Anticorpo secundário</td></tr><tr><td>Alexa Fluor 647 Goat anti-Rabbit</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-21244</td><td>Anticorpo secundário</td></tr><tr><td>Alexa Fluor 488 Goat anti-Mouse</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>A-11001</td><td>Anticorpo secundário</td></tr><tr><td>EVOS Microscope System</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Imageamento</td></tr><tr><td>TRI Reagent</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>T9424</td><td>Extração de RNA</td></tr><tr><td>Direct-zol RNA Kit</td><td>Zymo Research (Irvine, CA, USA)</td><td>R2050</td><td>Purificação de RNA</td></tr><tr><td>NanoDrop ND-1000</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>ND-1000</td><td>Quantificação de RNA</td></tr><tr><td>TaqMan Universal PCR Master Mix</td><td>Applied Biosystems (Foster City, CA, USA)</td><td>4304437</td><td>qPCR</td></tr><tr><td>QuantStudio 6 Flex Thermocycler</td><td>Life Technologies (Carlsbad, CA, USA)</td><td>N/A</td><td>qPCR</td></tr></tbody></table>

estabelecimento de culturas primárias enriquecidas em zona a partir do córtex adrenal do camundongo

Establishment of Zone-Enriched Primary Cultures from the Mouse Adrenal Cortex

Estabelecimento e caracterização de culturas celulares adrenais primárias enriquecidas em zonas Culturas primárias de células adrenais foram estabelecidas com sucesso a partir da OF, enriquecida para células zG, e da FI, enriquecida para células zF. Imagens representativas das células cultivadas são mostradas na Figura 2A (OF–zG) e na Figura 2B (IF–zF). Para avaliar o enriquecimento de células zG e não-zG nes...

Estabelecimento de Culturas Primárias Enriquecidas em Zona a partir do córtex adrenal do camundongo

The mouse adrenal gland is encapsulated by a mesenchymal cell layer (capsule) and contains an underlying cortex organized into distinct concentric zones ...

establishment-zone-enriched-primary-cultures-from-mouse-adrenal

Research

JoVE Journal

Biology

Biologia

Adrenal medular cromafins sistemas de cultura de células são extremamente úteis para o estudo da secreção de excitação de acoplamento em uma configuração em vitro. Este protocolo ilustra o método utilizado para dissecar as supra-renais e, em seguida, isolar a região medular, removendo o córtex adrenal. A digestão da medula em células cromafins único é então demonstrada.

Rato de isolamento de células Adrenal Cromafim

Este vídeo descreve o método utilizado para o isolamento de neuroprecursors desenvolvimento do córtex de ratos embrionárias. O procedimento para a remoção de embriões do útero, dissecando o tecido cortical, e digerindo o córtex cerebral isolado é mostrado.

Cultura de rato precursores das células-tronco neurais

Este artigo descreve um protocolo para isolamento e manutenção de culturas de fibroblastos primários da pele e do tecido pulmonar dos roedores silvestres.

Estabelecer culturas de fibroblastos adultos primária de roedores

Neste relatório nós descrevemos um método para o isolamento e cultura do nicho de células progenitoras do rim do rato embrionárias que podem ser usadas para estudar vias de sinalização que regulam tronco / progenitoras células do rim em desenvolvimento. Estas células em cultura são altamente acessível a pequenas moléculas e tratamento de proteínas recombinantes, e importante também para a transdução viral, que permite a manipulação eficiente das vias candidato.

Isolamento e cultura de células da zona nefrogênica do rim do rato embrionárias

Este artigo descreve métodos para derivar populações enriquecidas de células precursoras de oligodendrócitos murino (OPCs) em cultura primária, que se diferenciam para produzir oligodendrócitos maduros (MQO). Além disso, este relatório descreve as técnicas para produzir murino myelinating co-culturas semeando OPCs do mouse sobre uma cama de neurite do mouse neurônios do gânglio da raiz dorsal (DRGNs).

Derivação de Enriquecido Culturas oligodendroglial e Neuron / oligodendroglial Myelinating Co-culturas de tecidos pós-natais murino

Neste vídeo, descrevemos o método para obter uma preparação pura de astrócitos de camundongos recém-nascidos.

Isolamento de astrócitos: um método para obter preparação pura de astrócitos corticais de camundongos

Os astrócitos têm sido reconhecidos como células versáteis que participam em processos biológicos fundamentais, que são essenciais para o desenvolvimento do cérebro e função normais, e reparação do sistema nervoso central. Aqui apresenta-se um processo rápido de obtenção de culturas de astrócitos de rato puros para estudar a biologia desta classe importante de centrais células do sistema nervoso.

Isolamento e Cultura de astrócitos corticais de rato

Aqui, descrevemos um protocolo detalhado para a produção simultânea de culturas de células precursoras neurais, quer como monocamadas aderentes ou neuroesferas, a partir da zona subventricular e giro dentado de ratinhos adultos individuais.

Um rato, duas culturas: isolamento e cultivo de células-tronco neurais Adulto de as duas zonas neurogênica Individual Ratos

O epithelium retinal do pigment (RPE) é um epitélio multi-funcional do olho. Aqui nós apresentamos um protocolo para estabelecer culturas de célula primária derivadas o RPE murino.

Culturas de células primárias do Mouse epithelium Retinal do Pigment

Aqui nós apresentamos um método para isolar as glândulas adrenal de ratos, corrigir os tecidos, seção-los e executar a mancha da imunofluorescência.

Isolamento, fixação e imagem de imunofluorescência de glândulas supra-renais de Mouse

O hormônio secretado pelo córtex adrenal é vital para os animais contra doenças e estresse. Aqui, apresentamos um protocolo para cultura o rato primário células adrenais. Pode ser uma boa plataforma in-vitro para investigar os mecanismos do reagente de interesse na esteroidogênese adrenal e biossíntese de lipídios.

Cultura primária de células Adrenocortical rato e ensaios das funções reguladora

Aprenda a estabelecer culturas primárias enriquecidas em zona a partir do córtex adrenal do camundongo. Este método permite isolar células da zona glomerulosa e zona fasciculata para estudos de fisiologia adrenal.

Estabelecimento de Culturas Primárias Enriquecidas em Zona a partir do córtex adrenal do camundongo

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