Name: induction and analysis of oxidative stress in sleeping beauty transposon-transfected human retinal pigment epithelial cells
Uploaded: 2020-12-11T13:00:00
Description: Watch this Scientific Journal Video about Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells at JoVE.com

Forfatterne vil takke Gregg Sealy og Alain Conti for utmerket teknisk assistanse og prof. Zsuzsanna Izsvák fra Max-Delbrück Center i Berlin for vennlig å gi pSB100X og pT2-CAGGS-Venus plasmider. Dette arbeidet ble støttet av Swiss National Sciences Foundation og EU-kommisjonen i sammenheng med det syvende rammeprogrammet. Z.I ble finansiert av European Research Council, ERC Advanced [ERC-2011-ADG 294742].

Prosedyrer for innsamling og bruk av menneskelige øyne ble godkjent av Cantonal Ethical Commission for Research (nr. 2016-01726).
1. Celleisolasjon og kulturforhold
<ol><li>Menneskelig ARPE-19-cellelinje<ol>
<li>Kultur 5 x 105 ARPE-19 celler, en menneskelig RPE celle linje, i Dulbecco's Modified Eagle medium / næringsblanding F-12 Skin (DMEM / Hams F-12) supplert med 10% fetal bovine serum (FBS), 80 U / ml penicillin, 80 μg/ml streptomycin og 2,5 μg/ml amfotericin B (komplett ...

<ol>
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W., Van Houten, B., Conklin, C. A., Jin, G. F., Godley, B. 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Protokollen som presenteres her tilbyr en tilnærming for å analysere den antioksidative og beskyttende funksjonen til PEDF og GM-CSF produsert av transfected celler, som kan brukes på celler transfektert med ethvert putativt gunstig gen. I genterapeutiske strategier som har som mål å levere proteiner til vev ved å transplantere genetisk modifiserte celler, er det viktig å skaffe informasjon om nivået av proteinuttrykk, uttrykkslevelighet og effektiviteten av det uttrykte proteinet i en modell av sykdommen. I vår...

Modellen beskrevet her, tilbyr en nyttig tilnærming for å evaluere effektiviteten avbiofarmaceutiske midler for å redusere oksidativt stress i celler. Vi har brukt modellen til å undersøke de beskyttende effektene av PEDF og GM-CSF på H2O2-mediertoksidativt stress på retinal pigment epitelceller, som er utsatt for høye nivåer av O2, og synlig lys, og fagocytose av fotoreseptor ytre segmentmembraner, genererer betydelige nivåer av reaktive oksygenarter (ROS)1, 2. De regnes som en stor bidragsyter til patogenesen av avascular aldersrelatert makuladegenerasjon (aAMD)3,4,5,6,7,8. Dessuten er det en reduksjon i RPE-syntetiserte nevrobeskyttende faktorer, spesielt pigment epitel-avledet faktor (PEDF), insulin-lignende vekstfaktorer (IGFer), og granulocytt makrofag-kolonistimulerende faktor (GM-CSF) som fører til dysfunksjon og tap av RPE-celler, etterfulgt av fotoreseptor og retinal ganglion celle (RGC) død3,4 , 4 ,4, . AMD er en kompleks sykdom som skyldes samspillet mellom metabolske, funksjonelle, genetiske og miljømessige faktorer4. Mangelen på behandlinger for aAMD er hovedårsaken til blindhet hos pasienter eldre enn 60 år i industrialiserte land9,10. Rekonstitueringen av det nevrobeskyttende og nevrogene netthinnemiljøet ved subretinal transplantasjon av genetisk modifiserte RPE-celler som overekspresserer PEDF og GM-CSF har potensial til å forhindre retinal degenerasjon ved å redusere effekten av oksidativt stress, hemme betennelse og støtte celleoverlevelse11,12,13,14,15,16 . Selv om det er flere metoder for å levere gener til celler, har vi valgt det ikke-virale hyperaktive Sleeping Beauty transposon-systemet for å levere PEDF- og GM-CSF-genene til RPE-celler på grunn av sikkerhetsprofilen, integreringen av genene i vertscellenes genom, og dets tilbøyelighet til å integrere de leverte genene på ikke-transkripsjonsaktive steder som vi tidligere har vist17, 18,19.
Cellulært oksidativt stress kan induseres i celler dyrket in vitro av flere oksidative midler, inkludert hydrogenperoksid(H2O2), 4-hydroynonenal (HNE), tertbutylhydroperoxide (tBH), høye oksygenspenninger og synlig lys (fullspektret eller UV-bestråling)20,21. Høye oksygenspenninger og lys krever spesialutstyr og forhold, noe som begrenser overførbarheten til andre systemer. Agenter som H2O2, HNE og tBH induserer overlappende oksidativt stressmolekylære og cellulære endringer. Vi valgte H2O2 for å teste antioksidantaktiviteten til PEDF og GM-CSF fordi den er praktisk og biologisk relevant siden den produseres av RPE-celler som et reaktivt oksygen mellomliggende under fotoreseptor ytre segment fagocytose22 og det finnes i okulært vev in vivo23. Siden oksidasjonen av glutathione kan være delvis ansvarlig for produksjonen av H2O2 i øyet, har vi analysert nivåene av GSH / glutathione i våre studier, som er knyttet til H2O2-indusert oksidativt stress og den regenerative kapasiteten til cellene21,22. Analysen av glutathione nivåer er spesielt relevant siden den deltar i antioksidative beskyttelsesmekanismer i øyet24. Eksponering for H2O2 brukes ofte som modell for å undersøke oksidativ stressfølsomhet og antioksidantaktivitet i RPE-celler1,25,26,27,28,29,30, og i tillegg viser den likheter med lysindusert oksidativ stressskade, en "fysiologisk" kilde til oksidativt stress21.
For å evaluere funksjonaliteten og effektiviteten av nevrobeskyttende faktorer har vi utviklet en in vitro-modell som gjør det mulig for analysen å kvantifisere den antioksidative effekten av vekstfaktorer uttrykt av celler genetisk modifisert for å overekspressere PEDF og GM-CSF. Her viser vi at RPE-celler transfektert med genene for PEDF og GM-CSF er mer motstandsdyktige mot de skadelige effektene av H2O2 enn ikke-transfekerte kontrollceller, som det fremgår av cellemorfologi, tetthet, levedyktighet, intracellulært nivå av glutathione og uttrykk for UCP2-gen, som koder for mitokondrie-frakoblingsprotein 2 som har vist seg å redusere reaktive oksygenarter (ROS)31.

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			<td>24-well plates</td>
			<td>Corning</td>
			<td>353047</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well plates</td>
			<td>Greiner</td>
			<td>7657160</td>
			<td></td>
		</tr>
		<tr>
			<td>96-well culture plate white with clear flat bottom&#160;</td>
			<td>Costar</td>
			<td>3610</td>
			<td>Allows to check the cells before measuring the luminescence (GSH-Glo Assay)</td>
		</tr>
		<tr>
			<td>96-well plates</td>
			<td>Corning&#160;</td>
			<td>353072</td>
			<td></td>
		</tr>
		<tr>
			<td>Acrylamid 40%</td>
			<td>Biorad</td>
			<td>161-0144</td>
			<td></td>
		</tr>
		<tr>
			<td>Amphotericin B</td>
			<td>AMIMED</td>
			<td>4-05F00-H</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-GMCSF</td>
			<td>ThermoFisher Scientific</td>
			<td>PA5-24184</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-mouse IgG/IgA/IgM&#160;</td>
			<td>Agilent</td>
			<td>P0260</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-PEDF&#160;</td>
			<td>Santa Cruz Biotechnology Inc</td>
			<td>sc-390172</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-penta-His&#160;</td>
			<td>Qiagen</td>
			<td>34660</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-phospho-Akt</td>
			<td>Cell Signaling Technology</td>
			<td>9271</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody anti-rabbit IgG H&#38;L-HRP</td>
			<td>Abcam</td>
			<td>ab6721</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody donkey anti-rabbit Alexa Fluor 594&#160;</td>
			<td>ThermoFisher Scientific&#160;</td>
			<td>A11034</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibody goat anti-mouse Alexa 488</td>
			<td>ThermoFisher Scientific</td>
			<td>A-11029</td>
			<td></td>
		</tr>
		<tr>
			<td>ARPE-19 cell line</td>
			<td>ATCC</td>
			<td>CRL-2302</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>A9418-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>chamber culture glass slides</td>
			<td>Corning</td>
			<td>354118</td>
			<td></td>
		</tr>
		<tr>
			<td>CytoTox-Glo Cytotoxicity Assay&#160;</td>
			<td>Promega</td>
			<td>G9291</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Sigma-Aldrich</td>
			<td>D9542-5MG</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/Ham`s F12&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>D8062</td>
			<td></td>
		</tr>
		<tr>
			<td>Duo Set ELISA kit</td>
			<td>R&#38;D Systems&#160;</td>
			<td>DY215-05</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>ThermoFisher Scientific</td>
			<td>78440</td>
			<td></td>
		</tr>
		<tr>
			<td>ELISAquant kit</td>
			<td>BioProducts MD</td>
			<td>PED613-10-Human</td>
			<td></td>
		</tr>
		<tr>
			<td>Eyes (human)</td>
			<td>Lions Gift of Sight Eye Bank (Saint Paul, MN)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>FBS&#160;</td>
			<td>Brunschwig</td>
			<td>P40-37500</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluoromount Aqueous Mounting Medium</td>
			<td>Sigma-Aldrich</td>
			<td>F4680-25ML</td>
			<td></td>
		</tr>
		<tr>
			<td>FLUOstar Omega plate reader&#160;</td>
			<td>BMG Labtech</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>GraphPad Prism software (version 8.0)</td>
			<td>GraphPad Software, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>GSH-Glo Glutathione Assay</td>
			<td>Promega</td>
			<td>V6912</td>
			<td></td>
		</tr>
		<tr>
			<td>hydrogen peroxide (H2O2)</td>
			<td>Merck</td>
			<td>107209</td>
			<td></td>
		</tr>
		<tr>
			<td>ImageJ software (image processing program)</td>
			<td>W.S. Rasband, NIH, Bethesda, MD, USA; https://imagej.nih.gov/ij/; 1997&#8211;2014</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Imidazol&#160;</td>
			<td>Axonlab</td>
			<td>A1378.0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Leica DMI4000B microscope&#160;</td>
			<td>Leica Microsystems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LightCycler 480 Instrument II&#160;</td>
			<td>Roche Molecular Systems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LightCycler 480 SW1.5.1 software</td>
			<td>Roche Molecular Systems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma-Aldrich</td>
			<td>71376-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Axonlab</td>
			<td>3468.1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Neon Transfection System&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>MPK5000</td>
			<td></td>
		</tr>
		<tr>
			<td>Neon Transfection System 10 &#181;L Kit</td>
			<td>ThermoFisher Scientific</td>
			<td>MPK1096</td>
			<td></td>
		</tr>
		<tr>
			<td>Neubauer chamber</td>
			<td>Marienfeld-superior</td>
			<td>640010</td>
			<td></td>
		</tr>
		<tr>
			<td>Ni-NTA superflow&#160;</td>
			<td>Qiagen</td>
			<td>30410</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrocellulose&#160;</td>
			<td>VWR</td>
			<td>732-3197</td>
			<td></td>
		</tr>
		<tr>
			<td>Omega Lum G Gel Imaging System</td>
			<td>Aplegen Life Science</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PBS 1X</td>
			<td>Sigma-Aldrich</td>
			<td>D8537</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Sigma-Aldrich</td>
			<td>P0781-100</td>
			<td></td>
		</tr>
		<tr>
			<td>PerfeCTa SYBR Green FastMix</td>
			<td>Quantabio</td>
			<td>95072-012</td>
			<td></td>
		</tr>
		<tr>
			<td>PFA&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>158127-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Pierce BCA Protein Assay Kit&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>23227</td>
			<td></td>
		</tr>
		<tr>
			<td>Primers&#160;</td>
			<td>Invitrogen&#160;</td>
			<td></td>
			<td>See Table 1 in Supplementary Materials</td>
		</tr>
		<tr>
			<td>pSB100X (250 ng/&#181;L)</td>
			<td></td>
			<td></td>
			<td>M&#225;t&#233;s et al., 2009. Provide by Prof. Zsuzsanna Izsvak</td>
		</tr>
		<tr>
			<td>pT2-CMV-GMCSF-His plasmid DNA (250 ng/&#181;L)</td>
			<td></td>
			<td></td>
			<td>Constructed using the existing pT2-CMV-PEDF-EGFP plasmid reported in Johnen, S. et al. (2012) IOVS, 53 (8), 4787-4796.</td>
		</tr>
		<tr>
			<td>pT2-CMV-PEDF-His plasmid DNA (250 ng/&#181;L)</td>
			<td></td>
			<td></td>
			<td>Constructed using the existing pT2-CMV-PEDF-EGFP plasmid reported in Johnen, S. et al. (2012) IOVS, 53 (8), 4787-4796.</td>
		</tr>
		<tr>
			<td>QIAamp DNA Mini Kit</td>
			<td>QIAGEN</td>
			<td>51304</td>
			<td></td>
		</tr>
		<tr>
			<td>recombinant hGM-CSF&#160;</td>
			<td>Peprotech</td>
			<td>100-11</td>
			<td></td>
		</tr>
		<tr>
			<td>recombinant hPEDF&#160;&#160;</td>
			<td>BioProductsMD</td>
			<td>004-096</td>
			<td></td>
		</tr>
		<tr>
			<td>ReliaPrep RNA Cell Miniprep System</td>
			<td>Promega</td>
			<td>Z6011</td>
			<td></td>
		</tr>
		<tr>
			<td>RIPA buffer</td>
			<td>ThermoFisher Scientific</td>
			<td>89901</td>
			<td></td>
		</tr>
		<tr>
			<td>RNase-free DNase Set</td>
			<td>QIAGEN</td>
			<td>79254</td>
			<td></td>
		</tr>
		<tr>
			<td>RNeasy Mini Kit</td>
			<td>QIAGEN</td>
			<td>74204</td>
			<td></td>
		</tr>
		<tr>
			<td>SDS</td>
			<td>Applichem</td>
			<td>A2572</td>
			<td></td>
		</tr>
		<tr>
			<td>Semi-dry transfer system for WB&#160;</td>
			<td>Bio-Rad</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SuperMix qScript</td>
			<td>Quantabio</td>
			<td>95048-025</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-buffered saline (TBS)&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>15504020</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>AppliChem</td>
			<td>A4975</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin/EDTA</td>
			<td>Sigma-Aldrich</td>
			<td>T4174</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween</td>
			<td>AppliChem&#160;</td>
			<td>A1390</td>
			<td></td>
		</tr>
		<tr>
			<td>Urea</td>
			<td>ThermoFisher Scientific</td>
			<td>29700</td>
			<td></td>
		</tr>
		<tr>
			<td>WesternBright ECL HRP substrate</td>
			<td>Advansta</td>
			<td>K-12045-D50</td>
			<td></td>
		</tr>
		<tr>
			<td>Whatman nitrocellulose membrane</td>
			<td>Chemie Brunschwig</td>
			<td>MNSC04530301</td>
			<td></td>
		</tr>
	</tbody>
</table>

induction and analysis of oxidative stress in sleeping beauty transposon-transfected human retinal pigment epithelial cells

Oksidativt stress spiller en kritisk rolle i flere degenerative sykdommer, inkludert aldersrelatert makuladegenerasjon (AMD), en patologi som påvirker ~ 30 millioner pasienter over hele verden. Det fører til en reduksjon i retinal pigment epitel (RPE)-syntetisert neuroprotective faktorer, for eksempel pigment epitel-avledet faktor (PEDF) og granulocyte-makrophage kolonistimulerende faktor (GM-CSF), etterfulgt av tap av RPE celler, og til slutt fotoreseptor og retinal ganglion celle (RGC) død. Vi hypoteser at rekonstituering av nevrobeskyttende og nevrogen retinal miljø ved subretinal transplantasjon av transfected RPE celler overekspresserende PEDF og GM-CSF har potensial til å forhindre retinal degenerasjon ved å redusere effekten av oksidativt stress, hemme betennelse, og støtte celle overlevelse. Ved hjelp av Sleeping Beauty transposon-systemet (SB100X) har menneskelige RPE-celler blitt transfektet med PEDF- og GM-CSF-genene og vist stabil genintegrasjon, langsiktig genuttrykk og proteinsekresjon ved hjelp av qPCR, vestlig blot, ELISA og immunfluorescence. For å bekrefte funksjonaliteten og styrken til PEDF og GM-CSF utskilt av de transfekterte RPE-cellene, har vi utviklet en in vitro-analyse for å kvantifisere reduksjonen av H2O2-indusert oksidativt stress på RPE-celler i kultur. Cellebeskyttelse ble evaluert ved å analysere cellemorfologi, tetthet, intracellulært nivå av glutathione, UCP2 genuttrykk og celle levedyktighet. Begge, transfekerte RPE-celler som overekspresserte PEDF og/eller GM-CSF og celler som ikke var transfekterte, men forbehandlet med PEDF og/eller GM-CSF (kommersielt tilgjengelig eller renset fra transfekterte celler) viste betydelig antioksidantcellebeskyttelse sammenlignet med ikke-behandlede kontroller. Den nåværende H2O2-modellener en enkel og effektiv tilnærming for å evaluere antioksidanteffekten av faktorer som kan være effektive for å behandle AMD eller lignende nevrodegenerative sykdommer.

Induksjon av oksidativt stress i humane retinal pigment epitelceller ARPE-19 og primære hRPE-celler ble behandlet med varierende konsentrasjoner av H2O2 i 24 timer og det intracellulære nivået av antioksidantglutseksjonen ble kvantifisert (Figur 2A,B). H2O2 ved 50 μM og 100 μM påvirket ikke glutathioneproduksjonen, mens ved 350 μM var det en signifikant reduksjon av glutat...

Watch this Scientific Journal Video about Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells at JoVE.com

Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

Vi presenterer en protokoll for utvikling og bruk av en oksidativ stressmodell ved å behandle retinal pigment epitelceller med H2O2, analysere cellemorfologi, levedyktighet, tetthet, glutathione og UCP-2 nivå. Det er en nyttig modell for å undersøke antioksidanteffekten av proteiner utskilt av transposon-transfekerte celler for å behandle nevroretinal degenerasjon.

Induksjon og analyse av oksidativt stress i sovende skjønnhet transposon-transfekterte humane retinal pigment epitelceller

Oxidative stress plays a critical role in several degenerative diseases, including age-related macular degeneration (AMD), a pathology that affects ~30 ...

Research

JoVE Journal

Medicine

Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

Identification of Sleeping Beauty Transposon Insertions in Solid Tumors using Linker-mediated PCR

Identification of Sleeping Beauty Transposon Insertions in Solid Tumors using Linker-mediated PCR

Identifisering av<em&gt; Sleeping Beauty</em&gt; Transposon Innsettinger i solide svulster ved hjelp Linker-mediert PCR

Genomic, proteomic, transcriptomic, and epigenomic analyses of human tumors indicate that there are thousands of anomalies within each cancer genome ...

Transcriptomic Analysis of Human Retinal Surgical Specimens Using jouRNAl

Transcriptomic Analysis of Human Retinal Kirurgiske Prøver Bruke Journal

Retinal detachment (RD) describes a separation of the neurosensory retina  from the retinal pigmented epithelium (RPE). The RPE is essential for normal  ...

Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications

Isolasjon, Kryopreservering og kultur menneskerettighets amnion Epitelceller for kliniske anvendelser

Human amnion epithelial cells (hAECs) derived from term or pre-term amnion membranes have attracted attention from researchers and clinicians as a ...

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension

Utføre subretinal injeksjoner i Gnagere å levere retinal pigment epitelet Cells i Suspension

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Studerer Bukspyttkjertelkreft Stem Cell Kjennetegn for å utvikle nye behandlingsstrategier

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

A Step by Step Protocol for Subretinal Surgery in Rabbits

En trinnvis protokoll for subretinal Surgery i kaniner

Age related macular degeneration (AMD), retinitis pigmentosa, and other RPE related diseases are the most common causes for irreversible loss of vision in ...

Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

Nedbryting av museceller fra menneske tumorxenotransplantater forbedrer Nedstrøms Analyse av Target Cells

The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Induksjon av akselerert aterosklerose hos mus: "Wire-Injury" Modell

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Fosfor-31 Magnetic Resonance spektroskopi: Et verktøy for måling<em&gt; I Vivo</em&gt; Mitokondriell oksidativ fosforylering Kapasitet i Human Skeletal Muscle

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo ...