Name: Vestiging van zone-verrijkte primaire culturen uit de bijnierschors van de muis (Video) | JoVE Journal
Uploaded: 2026-05-08T14:20:00.000+00:00
Duration: 7 min 43 s
Description: Leer een fractioneringsprotocol voor zone-verrijkte primaire culturen uit de bijnierschors van de muis. Deze methode isoleert zona glomerulosa en zona fasciculata cellen voor gedetailleerd onderzoek naar hormoonproductie en signaalroutes.

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</ol>

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(Corning, NY, USA)</td><td>3527</td><td>Behandeld voor weefselcultuur</td></tr><tr><td>Steriele Filterpapier</td><td>N/A</td><td>N/A</td><td>Verwijdering van vet en ondersteuning bij dissectie</td></tr><tr><td>DMEM/F-12 Medium</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>11320033</td><td>1:1 formulering</td></tr><tr><td>Foetaal Rundserum (FBS)</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>10082147</td><td>Hitte-geïnactiveerd</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>Penicilline-Streptomycine (100X)</td><td>Gibco, Thermo Fisher Scientific (Waltham, MA, USA)</td><td>15140122</td><td>Antibioticum</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>Weefseldijging</td></tr><tr><td>DNase I</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>DN25-1</td><td>Weefseldijging</td></tr><tr><td>Angiotensine II (Ang II)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>A9525</td><td>Hormoonstimulatie</td></tr><tr><td>Adrenocorticotroop Hormoon (ACTH)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>A0298</td><td>Hormoonstimulatie</td></tr><tr><td>Forskolin</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>F6886</td><td>cAMP stimulatie</td></tr><tr><td>Kaliumchloride (KCl)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>P9541</td><td>Aldosteronstimulatie</td></tr><tr><td>Natriumchloride (NaCl)</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>S7653</td><td>Voertuigcontrole</td></tr><tr><td>Kleine chirurgische schaar</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>14060-09</td><td>Dissectie</td></tr><tr><td>Scalpel met wegwerpblad</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>10003-12</td><td>Microdissectie</td></tr><tr><td>Fijne pincet</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>11252-00</td><td>Dissectie</td></tr><tr><td>Gebogen pincet</td><td>Fine Science Tools (Foster City, CA, USA)</td><td>11251-10</td><td>Dissectie</td></tr><tr><td>Laminairstroomkap</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Steriele weefselverwerking</td></tr><tr><td>Incubator</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Celcultuur</td></tr><tr><td>Centrifuge</td><td>Eppendorf (Hamburg, Duitsland)</td><td>N/A</td><td>Celpelleting</td></tr><tr><td>Neubauer Tellingkamer</td><td>Hausser Scientific (Horsham, PA, USA)</td><td>1490</td><td>Celtelling</td></tr><tr><td>RIPA Lysis- en extractiebuffer</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>89901</td><td>Eiwitextractie</td></tr><tr><td>Halt Protease Inhibitor Cocktail</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>87786</td><td>Eiwitextractie</td></tr><tr><td>Fisherbrand Model 120 Sonic Dismembrator</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>FB120</td><td>Sonicatie</td></tr><tr><td>RIA Systeem</td><td>Tecan (M&auml;nnedorf, Zwitserland)</td><td>MG13051</td><td>Hormoonkwantificering</td></tr><tr><td>Cobra II Auto-Gamma Teller</td><td>Berthold Technologies (Bad Wildbad, Duitsland)</td><td>N/A</td><td>Radioactiviteitsmeting</td></tr><tr><td>Corticosterone ELISA Kit (96-well)</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>55-CORMS-E01</td><td>Hormoonmeting</td></tr><tr><td>ProLong Gold Antifade Mountant</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>P36930</td><td>Monteermiddel</td></tr><tr><td>Muis anti-Dab2</td><td>BD Biosciences (San Jose, CA, USA)</td><td>610464</td><td>Primaire antilichaam</td></tr><tr><td>Kip anti-GFP</td><td>Aves Labs (Tigard, OR, USA)</td><td>GFP-1020</td><td>Primaire antilichaam</td></tr><tr><td>Konijn anti-RFP</td><td>Rockland Immunochemicals (Limerick, PA, USA)</td><td>600-401-379</td><td>Primaire antilichaam</td></tr><tr><td>Alexa Fluor 594 Geit anti-Muis</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-11005</td><td>Secundair antilichaam</td></tr><tr><td>Alexa Fluor 594 Geit anti-Kip</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-11042</td><td>Secundair antilichaam</td></tr><tr><td>Alexa Fluor 647 Geit anti-Konijn</td><td>Invitrogen (Carlsbad, CA, USA)</td><td>A-21244</td><td>Secundair antilichaam</td></tr><tr><td>Alexa Fluor 488 Geit anti-Muis</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>A-11001</td><td>Secundair antilichaam</td></tr><tr><td>EVOS Microscoopsysteem</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>N/A</td><td>Beeldvorming</td></tr><tr><td>TRI Reagent</td><td>Sigma-Aldrich (St. Louis, MO, USA)</td><td>T9424</td><td>RNA-extractie</td></tr><tr><td>Direct-zol RNA Kit</td><td>Zymo Research (Irvine, CA, USA)</td><td>R2050</td><td>RNA-zuivering</td></tr><tr><td>NanoDrop ND-1000</td><td>Thermo Fisher Scientific (Waltham, MA, USA)</td><td>ND-1000</td><td>RNA-kwantificering</td></tr><tr><td>TaqMan Universal PCR Master Mix</td><td>Applied Biosystems (Foster City,

vestiging van zone-verrijkte primaire culturen uit de bijnierschors van de muis

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

Vestiging en karakterisering van zone-verrijkte primaire bijniercelculturen Primaire bijnierkweken werden succesvol gevestigd uit de OF, verrijkt voor zG-cellen, en de IF, verrijkt voor zF-cellen. Representatieve afbeeldingen van de gekweekte cellen zijn te zien in Figuur 2A (OF–zG) en Figuur 2B (IF–zF). Om de verrijking van zG-cellen en niet-zG-cellen in deze culturen te beoordelen, werd het ASCre/+</su...

Vestiging van zone-verrijkte primaire culturen uit de bijnierschors van de muis

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

Biologie

Bijniermerg chromaffin celkweek systemen zijn uiterst nuttig voor de studie van de excitatie-secretie koppeling in een in vitro setting. Dit protocol geeft de methode die wordt gebruikt om de bijnieren ontleden en vervolgens de medullaire regio te isoleren door het strippen van weg de bijnierschors. De vertering van de medulla in enkele chroomaffiene cellen wordt dan aangetoond.

Muis bijnier Chromaffinecellen Cell Isolatie

Deze video beschrijft de methode die wordt gebruikt voor het isoleren van neuroprecursors uit de ontwikkelingslanden cortex van embryonale muizen. De procedure voor het verwijderen van embryo's uit de baarmoeder, het ontleden van de corticale weefsel, en verteren de geïsoleerde cerebrale cortex wordt weergegeven.

Cultuur van Mouse Neurale Stem Cell Voorlopers

Dit artikel beschrijft een protocol voor de isolatie en het onderhoud van de primaire fibroblast culturen van de huid en longweefsel van wilde knaagdieren.

Tot vaststelling van primaire Adult Fibroblast Culturen van knaagdieren

In dit rapport beschrijven we een methode voor de isolatie en de cultuur van de progenitor cel niche van de embryonale muis nier die gebruikt kan worden om signaalwegen reguleren van stam / voorlopercellen van de zich ontwikkelende nier te bestuderen. Deze gekweekte cellen zijn zeer toegankelijk is voor kleine moleculen en recombinant eiwit behandeling, en vooral ook om virale transductie, die een efficiënte manipulatie van de kandidaat-wegen mogelijk maakt.

Isolatie en Cultuur van Cellen van de nefrogene Zone van de embryonale Mouse Kidney

Dit artikel beschrijft methoden voor het afleiden van verrijkte populaties van muizen oligodendrocyt voorloper cellen (OPC's) in het primair cultuur, die een onderscheid te rijpen oligodendrocyten (OLS) te produceren. Daarnaast is dit rapport beschrijft technieken te produceren voor muriene myelinating co-culturen door zaaien muis OPC's op een bedje van neurieten muis dorsale wortel ganglion neuronen (DRGNs).

Afleiding van Verrijkt oligodendrocyt Culturen en oligodendrocyt / Neuron Myelinating co-culturen van Post-natale Murine Tissues

In this video, we describe the method to obtain a pure preparation of astrocytes from newly born mice.

Astrocyte Isolation: A Method to Obtain Pure Preparation of Mouse Cortical Astrocytes

Astrocyten zijn erkend als veelzijdige cellen die deelnemen in de fundamentele biologische processen die essentieel zijn voor de normale ontwikkeling van de hersenen en functies binnen het centraal zenuwstelsel repareren. Hier presenteren we een snelle procedure voor pure muis astrocyten culturen te verkrijgen om de biologie van deze belangrijke klasse van het centrale zenuwstelsel cellen te bestuderen.

Isolatie en Cultuur van de Muis Corticale Astrocyten

Hier beschrijven we een gedetailleerd protocol voor het gelijktijdig opwekken van neurale precursoren celculturen, hetzij als hechtende monolagen of neurosferen, uit de subventriculaire zone en dentate gyrus van individuele volwassen muizen.

Een muis, twee culturen: Isolatie en Cultuur van adulte neurale stamcellen uit de Twee Neurogeen Zones van individuele muizen

De retinale pigment epitheel (RPE) is een multi-functionele epitheel van het oog. Hier presenteren we een protocol om primaire celculturen afgeleid van de lymfkliertest RPE.

Primaire celculturen van de muis retinale Pigment epitheel

Hier presenteren we een methode om te isoleren van de bijnieren van muizen, repareren de weefsels, sectie hen en uitvoeren immunofluorescentie kleuring.

Afzondering, fixatie en immunofluorescentie beeldvorming van muis bijnieren

Het hormoon afgescheiden door de bijnierschors is van vitaal belang voor dieren tegen stress en ziekten. Hier presenteren we een protocol bij de cultuur de primaire rat bijnier cellen. Het zou een goede in-vitro platform voor onderzoek naar de mechanismen van het reagens van belang in de bijnier steroidogenesis en lipide biosynthese.

Primaire cultuur van Rat Adrenocortical cellen en testen van Steroidogenic functies

Leer een fractioneringsprotocol voor zone-verrijkte primaire culturen uit de bijnierschors van de muis. Deze methode isoleert zona glomerulosa en zona fasciculata cellen voor gedetailleerd onderzoek naar hormoonproductie en signaalroutes.

Vestiging van zone-verrijkte primaire culturen uit de bijnierschors van de muis

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