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<ol>
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<table><tbody><tr><td>100 mm plate</td><td>SPL Life Science</td><td>10101SPL</td><td></td></tr><tr><td>10x Tris/Glycine Buffer</td><td>BioRad</td><td>1610734</td><td></td></tr><tr><td>12 mm glass coverslips</td><td>Marienfeld</td><td>111520</td><td></td></tr><tr><td>12-well plate</td><td>SPL Life Science</td><td>30012SPL</td><td></td></tr><tr><td>16% Paraformaldehyde Aqueous Solution</td><td>Invitrogen</td><td>28908</td><td></td></tr><tr><td>18 mm glass coverslips</td><td>HART</td><td>-</td><td></td></tr><tr><td>24-well plate</td><td>SPL Life Science</td><td>30024SPL</td><td></td></tr><tr><td>3,3',5-Triiodo-L-thyronine sodium salt (T3)</td><td>Sigma</td><td>T6397</td><td></td></tr><tr><td>3-Isobutyl-1-methylxanthine (IBMX)</td><td>Calbiochem</td><td>410957</td><td></td></tr><tr><td>30% Acrylamide/Bis Solution, 37.5:1</td><td>BioRad</td><td>1610158</td><td></td></tr><tr><td>35 mm plate</td><td>SPL Life Science</td><td>20035SPL</td><td></td></tr><tr><td>6-well plate</td><td>SPL Life Science</td><td>30006SPL</td><td></td></tr><tr><td>60 mm culture dish</td><td>SPL Life Science</td><td>20060SPL</td><td>Wordt gebruikt voor primaire preadipocyten en DNA/qPCR-experimenten; leverancier/catalogus niet verstrekt</td></tr><tr><td>AccuRuler RGB Plus Pre-stained Protein Ladder</td><td>Maestrogen</td><td>02102-250</td><td></td></tr><tr><td>ACK lysing buffer</td><td>Gibco</td><td>A10492-01</td><td></td></tr><tr><td>Actrapid 100 UI/mL (Insulin)</td><td>Novo Nordisk</td><td>A10AB01</td><td></td></tr><tr><td>ADP</td><td>Merck</td><td>117105</td><td></td></tr><tr><td>Agarose</td><td>FERMELO BIOTEC</td><td>FER00AL200G</td><td>Wordt gebruikt voor RNA-integriteitsverificatie; leverancier/catalogus niet verstrekt</td></tr><tr><td>Ammonium Persulfate</td><td>BioRad</td><td>1610700</td><td></td></tr><tr><td>Anti-rabbit IgG, HRP-linked Antibody</td><td>Cell Signaling</td><td>7074</td><td></td></tr><tr><td>Anti-rat IgG, HRP-linked Antibody</td><td>Cell Signaling</td><td>7077</td><td></td></tr><tr><td>Antibiotic-antimycotic</td><td>Gibco</td><td>15240062</td><td></td></tr><tr><td>Axio Observer inverted microscope stand</td><td>Carl Zeiss</td><td>431007-9901-000</td><td>Toegevoegd vanuit aanvullende methoden</td></tr><tr><td>BCA Protein Assay Kit</td><td>Thermo Scientific</td><td>D49328</td><td></td></tr><tr><td>BD Cytometer Setup and Tracking beads</td><td>BD Biosciences</td><td>-</td><td>Toegevoegd vanuit aanvullende methoden; catalogus niet verstrekt</td></tr><tr><td>BD FACSDiva Software v8.0.3</td><td>BD Biosciences</td><td>-</td><td>Toegevoegd vanuit aanvullende methoden</td></tr><tr><td>BD LSRFortessa X-20 cell analyzer</td><td>BD Biosciences</td><td>-</td><td>Bestaande invoer gestandaardiseerd om overeen te komen met aanvullend bestand</td></tr><tr><td>BioRender</td><td>BioRender</td><td>-</td><td>Wordt gebruikt om de workflow van Figuur 1 te maken</td></tr><tr><td>Blotting-grade blocker</td><td>BioRad</td><td>170-6404</td><td></td></tr><tr><td>BODIPY 493/503</td><td>Invitrogen</td><td>D3922</td><td></td></tr><tr><td>Bovine Serum Albumin (BSA)</td><td>Sigma</td><td>A1470</td><td></td></tr><tr><td>CaCl2</td><td>Calbiochem</td><td>208291</td><td></td></tr><tr><td>cDNA Reverse Transcription Kit</td><td>Invitrogen</td><td>4374966</td><td></td></tr><tr><td>Cell Strainer 100 μm, nylon</td><td>Falcon</td><td>352360</td><td></td></tr><tr><td>Cell Strainer 40 μm, nylon</td><td>Falcon</td><td>352340</td><td></td></tr><tr><td>Collagenase type II</td><td>Gibco</td><td>17101-015</td><td></td></tr><tr><td>Dexamethasone</td><td>Sigma</td><td>D4902</td><td></td></tr><tr><td>DMEM Powder, High Glucose, Pyruvate</td><td>Thermo Scientific</td><td>12800017</td><td></td></tr><tr><td>DMEM/F-12, no phenol red</td><td>Thermo Scientific</td><td>21041025</td><td></td></tr><tr><td>DMEM/F-12, powder</td><td>Gibco</td><td>12500062</td><td></td></tr><tr><td>DMSO</td><td>Sigma</td><td>D8418</td><td>Solvent voor inductiemedium; leverancier/catalogus niet verstrekt</td></tr><tr><td>EMBed-812 EMBEDDING KIT (Epon)</td><td>Electron Microscopy Sciences</td><td>14120</td><td></td></tr><tr><td>Ethanol absolute</td><td>Merck</td><td>100983</td><td></td></tr><tr><td>Ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid tetrasodium salt (EGTA)</td><td>Merck</td><td>324626</td><td></td></tr><tr><td>FAST SYBR MASTER MIX</td><td>Thermo Scientific</td><td>4385612</td><td></td></tr><tr><td>FCCP</td><td>Sigma</td><td>C2920</td><td></td></tr><tr><td>Fetal bovine serum</td><td>Capricorn Scientific</td><td>BS-16A</td><td></td></tr><tr><td>Floreada.io</td><td>Floreada.io</td><td>-</td><td>Flow cytometry analysesoftware</td></tr><tr><td>Fluoromount-G</td><td>Thermo Scientific</td><td>7984-25</td><td></td></tr><tr><td>GAPDH Antibody</td><td>Santa Cruz</td><td>sc-32233</td><td></td></tr><tr><td>Glucose</td><td>Gibco</td><td>15023-021</td><td></td></tr><tr><td>Glutamate</td><td>Sigma</td><td>G1626</td><td></td></tr><tr><td>Glutaraldehyde 25% Aqueous Solution</td><td>Electron Microscopy Sciences</td><td>16210</td><td></td></tr><tr><td>Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP</td><td>Invitrogen</td><td>62-6520</td><td></td></tr><tr><td>GraphPad Prism v10.6.1</td><td>GraphPad Software</td><td>-</td><td>Statistische/logaritmische regressieanalysesoftware</td></tr><tr><td>Halt Protease and Phosphatase Inhibitor Cocktail 100X</td><td>Thermo Scientific</td><td>78442</td><td></td></tr><tr><td>HEPES</td><td>Biological Industries</td><td>41-122-100</td><td></td></tr><tr><td>Histone H3 (1G1)</td><td>Santa Cruz</td><td>sc-517576</td><td></td></tr><tr><td>Hoechst 33342</td><td>Life Technologies</td><td>H3570</td><td></td></tr><tr><td>ImageJ v1.54s</td><td>NIH</td><td>-</td><td>Software voor analyse van lipidedruppelbeelden</td></tr><tr><td>Immun-Blot PVDF Membrane</td><td>BioRad</td><td>1620177</td><td></td></tr><tr><td>Indomethacin</td><td>Sigma</td><td>I7378</td><td></td></tr><tr><td>Insulin syringe</td><td>Cranberry</td><td>AAJECIN3</td><td>Wordt gebruikt voor celhomogenisatie; leverancier/catalogus niet verstrekt</td></tr><tr><td>KCl</td><td>Calbiochem</td><td>529552</td><td></td></tr><tr><td>KH2PO4</td><td>Calbiochem</td><td>529568</td><td></td></tr><tr><td>KHCO3</td><td>Sigma</td><td>60339</td><td></td></tr><tr><td>LAMP-1 Antibody</td><td>Abcam</td><td>ab25245</td><td></td></tr><tr><td>Lane Marker Reducing Sample Buffer</td><td>Thermo Scientific</td><td>39000</td><td></td></tr><tr><td>Malate</td><td>Sigma</td><td>M1000</td><td></td></tr><tr><td>MgCl2</td><td>Calbiochem</td><td>208291</td><td>On

exogene mitochondriale overdracht bij het differentiëren van bruine adipocyten en agpat2-deficiënte preadipocyten

Exogenous Mitochondrial Transfer in Differentiating Brown Adipocytes and AGPAT2-Deficient Preadipocytes

Exogene mitochondriale overdracht bij het differentiëren van bruine adipocyten en AGPAT2-deficiënte preadipocyten

Mitochondria are key signaling hubs; however, whether mitochondrial mass expansion is mechanistically required for differentiation remains an open ...

exogenous-mitochondrial-transfer-differentiating-brown-adipocytes

Research

JoVE Journal

Biology

Biologie

Primaire witte preadipocyten geïsoleerd van wit vetweefsel in muizen kunnen worden onderscheiden in beige / brite cellen. Hier wordt gepresenteerd is een betrouwbare cellulaire modelsysteem om de moleculaire regulatie van "bruinen" van wit vet te bestuderen.

Isolatie en differentiatie van Stroma vasculaire cellen te Beige / Brite Cellen

Hier, wordt een protocol (MitoCeption) voorgelegd aan mitochondriën geïsoleerd uit menselijke mesenchymale stamcellen (MSC) over te dragen, glioblastoma stamcellen (GSC), met als doel het bestuderen van hun biologische effecten op het metabolisme en de functies SGR. Een soortgelijk protocol kan worden aangepast aan mitochondriën dragen tussen andere celtypen.

MitoCeption: overbrengen geïsoleerde menselijke MSC Mitochondriën aan glioblastoma Stamcellen

Dit rapport beschrijft een protocol voor de gelijktijdige isolatie van primaire bruine en witte preadipocyten van pasgeboren muizen. Geïsoleerde cellen kunnen in cultuur worden gekweekt en worden geïnduceerd om zich te onderscheiden in volgroeide witte en bruine adipocyten. De methode maakt genetische, moleculaire en functionele karakterisering van primaire vetcellen in cultuur mogelijk.

Isolatie en differentiatie van primaire witte en bruine preadipocyten van pasgeboren muizen

Deze studie beschrijft een nieuwe methode voor het isoleren van muizenbruine adipocyten voor gen- en eiwitexpressieanalyse.

Het isoleren van bruine adipocyten uit murine interscapulair bruin vetweefsel voor gen- en eiwitexpressieanalyse

Functionele beeldvorming en kwantificering van thermogene vetdepots bij muizen met behulp van een micro-PET / MR-beeldvormingsgebaseerde benadering.

Visualisatie en kwantificering van bruine en beige vetweefsels bij muizen met behulp van [18F] FDG Micro-PET / MR Imaging

Preadipocyten worden geïsoleerd uit de stromale vasculaire fractie van interscapulier bruin vetweefsel van pasgeboren muizen en gedifferentieerd tot cellen die lipidedruppeltjes accumuleren, moleculaire markers tot expressie brengen en de mitochondriale morfologie van volwassen bruine vetcellen vertonen. Deze cellen worden verder geanalyseerd door middel van immunofluorescentie en transmissie-elektronenmicroscopie.

Differentiatie en beeldvorming van bruine vetcellen uit de stromale vasculaire fractie van interscapulier vetweefsel van pasgeboren muizen

Dit protocol beschrijft de semi-geautomatiseerde isolatie van de stromale vasculaire fractie (SVF) uit muizenvetweefsel om preadipocyten te verkrijgen en adipocytendifferentiatie in vitro te bereiken. Het gebruik van een weefseldissociator voor collagenasevertering vermindert experimentele variatie en verhoogt de reproduceerbaarheid.

Semi-geautomatiseerde isolatie van de stromale vasculaire fractie uit muizenwit vetweefsel met behulp van een weefseldissociator

Genetische Induceerbare Fate Mapping (GIFM) merken en tracks cellen met fijne ruimtelijke en temporele...

Een praktische benadering van genetische Induceerbare Fate Mapping: een visuele gids voor Mark en Track Cellen In Vivo</em

Het visualiseren van RNA Lokalisatie in Xenopus Eicellen

Hier beschrijven we protocollen om endogene en exogene centromeer-kinetochoor-eiwitten in menselijke cellen te detecteren en deze eiwitniveaus op centromeren-kinetochoren te kwantificeren door indirecte immunofluorescente kleuring door het gebruik van fixatie (paraformaldehyde, aceton of methanol fixatie).

Immunofluorescentie analyse van endogene en exogene Centromere-kinetochoor Eiwitten

Pontificia Universidad Católica de Chile.  Exogene mitochondriën van menselijke en muisachtige oorsprong kunnen worden geïnternaliseerd door preadipocyten te differentiëren en gedurende de adipogene differentiatie worden gevolgd met behulp van beeldvorming en genetische hulpmiddelen. Hoewel deze mitochondriën fysiek interageren met het endogene mitochondriale netwerk, verhogen ze de expressie van markers die geassocieerd zijn met volwassen bruine adipocyten niet.

Exogene mitochondriale overdracht bij het differentiëren van bruine adipocyten en AGPAT2-deficiënte preadipocyten

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