Name: Stabilimento di colture primarie arricchite in zona dalla corteccia surrenale del topo (Video) | JoVE Journal
Uploaded: 2026-05-08T14:20:00.000+00:00
Duration: 7 min 43 s
Description: Scopri come stabilire colture primarie arricchite in zona dalla corteccia surrenale del topo. Questo protocollo consente di isolare cellule della zona glomerulosa e fascicolata per studi di endocrinologia e fisiologia surrenale.

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</ol>

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Plate</td><td>Corning (Corning, NY, USA)</td><td>3527</td><td>Trattati per la cultura dei tessuti</td></tr><tr><td>Sterile Filter Paper</td><td>N/A</td><td>N/A</td><td>Rimozione del grasso e supporto per la dissezione</td></tr><tr><td>DMEM/F-12 Medium</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>11320033</td><td>Formulazione 1:1</td></tr><tr><td>Fetal Bovine Serum (FBS)</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>10082147</td><td>Heat-inactivated</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>Antibiotico</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>Digestione dei tessuti</td></tr><tr><td>DNase I</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>DN25-1</td><td>Digestione dei tessuti</td></tr><tr><td>Angiotensin II (Ang II)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>A9525</td><td>Stimolazione ormonale</td></tr><tr><td>Adrenocorticotropic Hormone (ACTH)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>A0298</td><td>Stimolazione ormonale</td></tr><tr><td>Forskolin</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>F6886</td><td>Stimolazione cAMP</td></tr><tr><td>Potassium Chloride (KCl)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>P9541</td><td>Stimolazione aldosterone</td></tr><tr><td>Sodium Chloride (NaCl)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>S7653</td><td>Controllo del veicolo</td></tr><tr><td>Small Surgical Scissors</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>14060-09</td><td>Dissezione</td></tr><tr><td>Scalpel with Disposable Blade</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>10003-12</td><td>Microdissezione</td></tr><tr><td>Fine Forceps</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>11252-00</td><td>Dissezione</td></tr><tr><td>Curved Forceps</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>11251-10</td><td>Dissezione</td></tr><tr><td>Laminar Flow Hood</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Elaborazione sterile del tessuto</td></tr><tr><td>Incubator</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Coltura cellulare</td></tr><tr><td>Centrifuge</td><td>Eppendorf (Hamburg, Germany)</td><td>N/A</td><td>Pellettizzazione delle cellule</td></tr><tr><td>Neubauer Counting Chamber</td><td>Hausser Scientific (Horsham, PA, USA)</td><td>1490</td><td>Conteggio delle cellule</td></tr><tr><td>RIPA Lysis and Extraction Buffer</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>89901</td><td>Estrazione delle proteine</td></tr><tr><td>Halt Protease Inhibitor Cocktail</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>87786</td><td>Estrazione delle proteine</td></tr><tr><td>Fisherbrand Model 120 Sonic Dismembrator</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>FB120</td><td>Sonicazione</td></tr><tr><td>RIA System</td><td>Tecan (M&auml;nnedorf, Switzerland)</td><td>MG13051</td><td>Quantificazione degli ormoni</td></tr><tr><td>Cobra II Auto-Gamma Counter</td><td>Berthold Technologies (Bad Wildbad, Germany)</td><td>N/A</td><td>Misurazione della radioattività</td></tr><tr><td>Corticosterone ELISA Kit (96-well)</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>55-CORMS-E01</td><td>Misurazione degli ormoni</td></tr><tr><td>ProLong Gold Antifade Mountant</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>P36930</td><td>Medium di montaggio</td></tr><tr><td>Mouse anti-Dab2</td><td>BD Biosciences (San Jose, CA, USA)</td><td>610464</td><td>Anticorpo primario</td></tr><tr><td>Chicken anti-GFP</td><td>Aves Labs (Tigard, OR, USA)</td><td>GFP-1020</td><td>Anticorpo primario</td></tr><tr><td>Rabbit anti-RFP</td><td>Rockland Immunochemicals (Limerick, PA, USA)</td><td>600-401-379</td><td>Anticorpo primario</td></tr><tr><td>Alexa Fluor 594 Goat anti-Mouse</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-11005</td><td>Anticorpo secondario</td></tr><tr><td>Alexa Fluor 594 Goat anti-Chicken</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-11042</td><td>Anticorpo secondario</td></tr><tr><td>Alexa Fluor 647 Goat anti-Rabbit</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-21244</td><td>Anticorpo secondario</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 secondario</td></tr><tr><td>EVOS Microscope System</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Imaging</td></tr><tr><td>TRI Reagent</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>T9424</td><td>Estrazione dell'RNA</td></tr><tr><td>Direct-zol RNA Kit</td><td>Zymo Research (Irvine, CA, USA)</td><td>R2050</td><td>Purificazione dell'RNA</td></tr><tr><td>NanoDrop ND-1000</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>ND-1000</td><td>Quantificazione dell'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>

stabilimento di colture primarie arricchite in zona dalla corteccia surrenale del topo

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

Stabilimento e caratterizzazione di colture cellulari surrenali primarie arricchite in zone Colture primarie di cellule surrenali sono state stabilite con successo dall'OF, arricchite per le cellule zG, e dall'IF, arricchite per le cellule zF. Immagini rappresentative delle cellule coltivate sono mostrate nella Figura 2A (OF–zG) e nella Figura 2B (IF–zF). Per valutare l'arricchimento delle cellule zG e non zG...

Stabilimento di colture primarie arricchite in zona dalla corteccia surrenale del topo

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

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JoVE Journal

Biology

Biologia

Midollare surrenale cromaffini sistemi di colture cellulari sono estremamente utili per lo studio della secrezione di accoppiamento eccitazione-in un ambiente in vitro. Questo protocollo illustra il metodo utilizzato per analizzare le ghiandole surrenali e quindi isolare la regione midollare strappando via la corteccia surrenale. La digestione del midollo in singole cellule cromaffini è poi dimostrato.

Topo surrenale cromaffini cella di isolamento

Questo video descrive il metodo utilizzato per l'isolamento di neuroprecursors dalla corteccia sviluppo embrionale dei topi. Il procedimento per eliminare gli embrioni dall'utero, sezionare il tessuto corticale, e digerire la corteccia cerebrale isolato viene mostrato.

Cultura di cellule precursori staminali neurali del mouse

Questo articolo descrive un protocollo per l'isolamento e il mantenimento delle colture primarie di fibroblasti della pelle e tessuto polmonare dei roditori selvatici.

Stabilire colture primarie di fibroblasti adulti dai roditori

In questo rapporto si descrive un metodo per l'isolamento e la coltura della nicchia delle cellule progenitrici dal rene embrionali di topo che può essere utilizzato per studiare vie di segnalazione che regolano staminali / progenitrici delle cellule del rene di sviluppo. Queste cellule in coltura sono altamente accessibile a piccole molecole e proteine ​​ricombinanti trattamento, e, cosa importante anche per trasduzione virale, che consente la manipolazione efficiente di percorsi candidato.

Isolamento e coltura di cellule dalla Zona nefrogenica del Rene embrionali di topo

Questo articolo descrive i metodi per ricavare le popolazioni murine arricchito di cellule precursori degli oligodendrociti (OPC) in colture primarie, che differenziano per produrre oligodendrociti maturi (OLS). Inoltre, questo rapporto descrive le tecniche per produrre murino mielinizzanti co-colture da semina OPC del mouse su un letto neurite dei neuroni dei gangli spinali del mouse (DRGNs).

Derivazione di Arricchito Culture oligodendrociti e degli oligodendrociti / Neuron mielinizzanti co-culture di tessuti murini post-natale

In questo video, descriviamo il metodo per ottenere una preparazione pura di astrociti da topi appena nati.

Isolamento degli astrociti: un metodo per ottenere una preparazione pura di astrociti corticali di topo

Gli astrociti sono stati riconosciuti per essere versatili cellule che partecipano a processi biologici fondamentali che sono essenziali per il normale sviluppo del cervello e la funzione, e la riparazione del sistema nervoso centrale. Qui vi presentiamo una procedura rapida per ottenere colture pure astrociti topo per studiare la biologia di questa classe importante di cellule del sistema nervoso centrale.

Isolamento e la coltura di astrociti corticali mouse

Qui si descrive un protocollo dettagliato per la generazione simultanea di colture di cellule precursore neuronali, sia come monostrati aderenti o neurosfere, dalla zona subventricolare e giro dentato di singoli topi adulti.

Un mouse, due culture: isolamento e la coltura di Adult Neural Stem Cells dalle due zone neurogena Mice individuale

L'epitelio pigmentato retinico (RPE) è un multi-funzionale dell'epitelio dell'occhio. Qui presentiamo un protocollo standard per stabilire colture cellulari primarie derivate da RPE murino.

Colture cellulari primarie dall'epitelio retinico del pigmento del Mouse

Qui presentiamo un metodo per isolare ghiandole surrenali da topi, difficoltà i tessuti, sezione loro ed eseguire la colorazione di immunofluorescenza.

Isolamento, fissazione e immunofluorescenza Imaging delle ghiandole adrenali Mouse

L'ormone secernuta dalla corteccia surrenale è vitale per animali contro lo stress e malattie. Qui, presentiamo un protocollo alla cultura il ratto primario cellule adrenali. Potrebbe essere una buona piattaforma in vitro per indagare i meccanismi del reagente di interesse nella steroidogenesi surrenalica e biosintesi lipidica.

Colture primarie di cellule corticosurrenali ratto e saggi di Steroidogenic funzioni

Scopri come stabilire colture primarie arricchite in zona dalla corteccia surrenale del topo. Questo protocollo consente di isolare cellule della zona glomerulosa e fascicolata per studi di endocrinologia e fisiologia surrenale.

Stabilimento di colture primarie arricchite in zona dalla corteccia surrenale del topo

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