Name: a high-content in vitro pancreatic islet &#946;-cell replication discovery platform
Uploaded: 2016-07-16T15:00:00
Description: Watch this Scientific Journal Video about A High-content In Vitro Pancreatic Islet ÃŽÂ²-cell Replication Discovery Platform at JoVE.com

This work was supported by NIDDK grants DK098143 and DK101530 from the NIH (JPA), Stanford's Spectrum Child Health Research Institute (CHRI) and SPARK (UL1 TR001085, JPA).

Denne protokollen ble utført i samsvar med Institutional Animal Care og bruk Committee (IACUC) ved Stanford University School of Medicine. Den beskrevne protokoll er skalert for holme isolert fra seks 250-300 g (8 - 9 uker gamle) Sprague Dawley hannrotter, som er tilstrekkelig til å generere 228 brønner i en 384-brønners plate for islet celle-replikasjon vurdering. 1. Material Forberedelse <ol><li> Forbered belegg media før start holme isolasjon ved å samle det kondisjonerte medium av 804G rotte...

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Eksperimentelle metoder for å studere de molekylære stier som styrer β-cellevekst og regenerering er viktige verktøy for diabetes forskere. Heri, til en rotte-islet-basert screening plattform identifisere og karakterisere små-molekyl stimulatorer for β-celle-replikasjon er presentert. Mens de fleste aspekter av denne protokollen er lett utføres av erfarne forskere, noen få skritt krever spesiell teknikk. Først i løpet av holmen isolasjon, kanylering av galle-kanal uten å forstyrre...

Diabetes omfatter en samling av forstyrrelser som deler den felles sluttpunkt forstyrret glukose homeostase. Selv om de patogene mekanismene for diabetes subtyper er adskilte, men de deler som følge av redusert β-cellemasse, dvs. tap av insulinproduksjonskapasitet 1,2. I dag, diabetes behandlingsstrategier stole på kronisk administrering av eksogent insulin, farmakologisk stimulering av insulin produksjon eller forbedring av insulinfølsomhet, og sjelden, transplantasjon av pankreas holmer eller hele bukspyttkjertelen 3,4. Dessverre, suksessen av disse strategiene er kortvarig og / eller unnlater å tilstrekkelig rekapitulere funksjonen av endogen insulinproduksjon. Til tross for anvendeligheten av å utvikle en metode for å stimulere β-celle regenerering, foreligger ingen slik tilnærming. Følgelig er et viktig diabetesforskning mål å utvikle metoder for å generere nye p-celler, eller for å utvide endogen β-cellemasse 5 </sup&gt;. Selv β-celle gjenfødelse fra fornybare kilder som embryonale stamceller er fremme, sikkerhets- og effektivitetshensyn gjør jakten på alternative strategier, herunder utvidelse av modne p-celler, en prioritet 6,7. Viktigere er den dominerende kilde til nye p-celler in vivo pre-eksisterende p-celler i stedet for spesialiserte stamceller 8,9. Selv om p-celler synes å ha begrenset kapasitet replikasjon, en liten økning i β-cellemasse (~ 30%) kan være tilstrekkelig til å gjenopprette glukosehomeostase i mange diabetikere. Videre in situ farmakologisk stimulering av β-cellemasse er et potensielt billig og skalerbar behandlingsstrategi. Heri et høyt innhold screening fremgangsmåte for å identifisere og karakterisere små molekyler som stimulerer β-cellevekst er presentert. En rekke in vitro eksperimentelle metoder kan anvendes for å identifisere genprodukter og / eller molekyler that fremme primære β-celle replikering. Tidlig innsats for måling av β-celle replikering induksjon brukes fosterets gnager pankreata kultur eller intakte isolert holmen kulturer å måle [3H] tymidin inkorporering, BrdU inkorporering eller mitotiske organer innenfor aldehyd-thionin eller insulin farget befolkning som svar på konkrete behandlingsforhold 10, 11. Disse in vitro tilnærminger og nære varianter av disse har flere begrensninger. Prominente mangler inkluderer (1) bruk av føtale celler som, i motsetning til modne p-celler, viser en høy basal β-celle replikering rate og er vekst regulert i en tydelig måte 12; (2) den subjektive natur eksperimentator avhengig pådømmelse av β-celle replikering hendelser; (3) arbeids- og tidkrevende natur eksperimentator avhengig telling av β-celle replikering hendelser forsinker eksperimentell gjennomstrømming; (4) bruk av kjernefysisk innlemmelse / beis / utseende til å identifisere replikering selvts og en ikke-overlappende cytoplasmisk flekken for å identifisere p-celler fører til misattribution av tilgrensende ikke-β-cellereplikasjonshendelser til p-celler. Mer nylig modne primære p-celler er blitt anvendt for å vurdere effekten av transgene over-ekspresjon, så vel som genprodukt eller behandling med forbindelsen på β-celle-replikasjon 13-16. Imidlertid har disse studiene også grunnlag subjektive telling av replikering hendelser, cytoplasmatiske staining- eller ikke-spesifikke metoder for β-celle identifisering og / eller arbeidskrevende tiltak som begrenser gjennomstrømning, f.eks enkelte slide-vel plating av celler eller intakt holme parafin embedding og behandling 17. Spesielt, en bildebasert human β-celle-replikasjon screening metode, lik den som er presentert heri, er blitt publisert 18; har imidlertid vellykket bruk av denne analysen ikke blitt demonstrert og bruken av humane øyer for primær screening kan ikke bli så å si fealig. En alternativ strategi for å identifisere replikasjonsfremmende stoffer er å vurdere veksten induksjon av β-cellelinjer. Innledende forsøk brukes forvandlet betacellelinjer som min6 celler eller INS 832/13-celler 14,19-21. Men disse cellelinjer demonstrere uhemmet vekst og har liten likhet med godt differensierte p-celler 22. Følgelig er minimalt, av uklare relevans og noen ganger vanskelig å rekapitulere vekst-induksjon kapasitet. En forbedret strategi for cellelinje basert screening benytter &quot;reversibelt transformerte&quot; celler som er veksten arrestert i fravær av tetracyklin (doksycyklin) -avhengig SV40 T-antigen ekspresjon 23,24. Imidlertid er det uklart om disse cellene gå tilbake til en &quot;normal&quot; β-celle-lignende tilstand etter doksycyklin fjerning. Dessverre har bruken av disse cellene ga en generell vekstfremmende stoffer som ikke synes å ha umiddelbar nytte24. Samlet bruk av cellelinjene for å studere vekstregulering av en celletype som viser minimal spontan replikasjonsaktivitet kan ha begrenset anvendelighet. Den β-celle replikering screening plattform som presenteres her benytter modne primære rotte p-celler til å beholde in vivo vekstregulering i den grad det er mulig, holme-cellekulturer med blandet celletype sammensetning for å muliggjøre identifisering av avstamning begrensede vekstfremmende aktiviteter, multi -Vel formatering for å maksimere gjennomstrømningen og automatisert analyse for å eliminere skjevhet og legge til rette for gjennomstrømming. Vellykket bruk av denne plattformen har gjort det mulig identifisering av flere forbindelser som fremmer β-celle replikering 25,26. I tillegg har analysen vært brukt til struktur-aktivitetsforhold studier og kjemiske epistasis eksperimenter for å tilveiebringe mekanistiske innsikt i den molekylære regulering av β-celle-replikasjon. Den presenterte plattformen ble vellykket tilpasset for lentiviral RNAi-basert undersøkelse av β-celle replikering trasé 25. Begrensninger av analysen omfatter bundne skalerbarhet (bruk av primærceller), bruk av gnager heller enn menneskelige holmen-celler (selv om analysen kan tilpasses for menneske holmen studier), kostnader forbundet med antistoff-basert bildebehandling og primær holme bruk, bruk av spredte holmer (forstyrret holme arkitektur) for å lette automatisert bilde oppkjøpet og avhengighet av tilgjengeligheten av en automatisert mikroskop med bilde oppkjøpet og analyse evne. Selv om en lettvint in vivo screening-metoder for identifisering av genprodukter eller forbindelser som stimulerer β-celle regenerering in situ ville være ideelt, er en slik plattform ikke ennå er tilgjengelig 27. Følgelig er hensiktsmessig for forskeren er interessert i å undersøke de fleste aspekter av β-celle replikering den beskrevne plattformen.

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			<td height="23" style="height:23px;width:364px;">250g male Male Sprague Dawly Rat</td>
			<td style="width:196px;">Charles River</td>
			<td style="width:147px;">Stain # 400</td>
			<td style="width:272px;"></td>
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			<td height="22" style="height:22px;width:364px;">12 cm teeth tisuue forceps</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">11021-12</td>
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			<td height="21" style="height:21px;width:364px;">11.5 cm fine scissors</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">14058-11</td>
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			<td height="21" style="height:21px;width:364px;">14.5 cm surgical scissors</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">14001-14</td>
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			<td height="21" style="height:21px;width:364px;">16 cm curved forceps</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">11003-16</td>
			<td></td>
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			<td height="21" style="height:21px;width:364px;">12 cm curved hepostat</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">13011-12</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">12 cm scalpel handle</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">10003-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Tissue sieve-30 mesh</td>
			<td style="width:196px;">Bellco Glass</td>
			<td style="width:147px;">1985-85000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Cizyme RI, 375,000 CDA units</td>
			<td style="width:196px;">VitaCyte</td>
			<td style="width:147px;">005-1030</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:364px;">Hanks&#39; Balanced Salt solution (Ca++ and Mg++)</td>
			<td style="width:196px;">Gibco</td>
			<td style="width:147px;">24020-117</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Ketamine HCl (200 mg/20 ml)</td>
			<td style="width:196px;">JHP Pharmaceuticals</td>
			<td style="width:147px;">NDC# 42023-113-10</td>
			<td>to make anesthetic cocktail&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Xylazine (5 g/50 ml)</td>
			<td style="width:196px;">LLOYD</td>
			<td style="width:147px;">NADA# 139-236</td>
			<td>to make anesthetic cocktail&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Histopaque 1077</td>
			<td style="width:196px;">Sigma</td>
			<td style="width:147px;">H-1077</td>
			<td>to make histopaque 1100</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Histopaque 1119</td>
			<td style="width:196px;">Sigma</td>
			<td style="width:147px;">H-1119</td>
			<td>to make histopaque 1100</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Newborn Calf Serum 500 ml</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30118.03</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Hanks&#39; Balanced Salt solution</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30268.01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dulbecco&#39;s Modified Eagle Medium/Low Glucose&#160;</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30021.01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Functionality/Viability Solution&#160;</td>
			<td style="width:196px;">Mediatech</td>
			<td style="width:147px;">99-768-CV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">RPMI1640 media&#160;</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30096.01</td>
			<td>to make conditioned medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">804G rat bladder carcinoma cell-line</td>
			<td style="width:196px;">Available upon request</td>
			<td style="width:147px;"></td>
			<td>to make conditioned medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Fetal Bovine Serum, Qualified</td>
			<td style="width:196px;">Gibco</td>
			<td style="width:147px;">26160</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">GlutaMax-I</td>
			<td style="width:196px;">Gibco</td>
			<td style="width:147px;">35050-061</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Penicillin (5,000 IU/ml/Strptomycin (5 mg/ml)&#160;</td>
			<td style="width:196px;">MP Biomedicals</td>
			<td style="width:147px;">1670049</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Formamide 500 mL</td>
			<td style="width:196px;">Fisher BioReagents</td>
			<td style="width:147px;">BP227-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Antigen Unmasking Solution 250 mL (PH 6.0)</td>
			<td style="width:196px;">Vector Laboratories</td>
			<td style="width:147px;">H-3300</td>
			<td>to make 0.15 M Sodium Sitrate solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Dextrose, Anhydrous</td>
			<td style="width:196px;">EMD Chemicals</td>
			<td style="width:147px;">DX0145-1</td>
			<td>to make 1 M glucose solution</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:364px;">Nomal Donkey Serum (Powder)</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">017-000-121</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Triton X-100</td>
			<td style="width:196px;">Sigma</td>
			<td style="width:147px;">T8787-100ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Mouse anti-human Ki67 antibody</td>
			<td style="width:196px;">BD Biosciences</td>
			<td style="width:147px;">556003</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Goat anti-human PDX-1 antibody</td>
			<td style="width:196px;">R&#38;D Systems</td>
			<td style="width:147px;">AF2419</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal Guinea Pig anti-insulin antibody</td>
			<td style="width:196px;">Dako</td>
			<td style="width:147px;">2016-08</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal Rabbit anti-glucagon antibody</td>
			<td style="width:196px;">Dako</td>
			<td style="width:147px;">2014-06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal Rabbit anti-somatostatin antibody</td>
			<td style="width:196px;">Dako</td>
			<td style="width:147px;">2011-08</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal chicken anti-vimentin antibody</td>
			<td style="width:196px;">abcam</td>
			<td style="width:147px;">ab24525</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biotin-SP-conjugated, Donkey Anti-Mouse IgG</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">715-065-150</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">StreptAvidin, Alex Flour 488 conjugated&#160;</td>
			<td style="width:196px;">Invitrogen</td>
			<td style="width:147px;">S32354</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Rhodamine-conjugated Donkey Anti-Goat IgG&#160;</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">705-025-147</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Rhodamine-conjugated Donkey Anti-Guinea Pig IgG&#160;</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">706-025-148</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Rhodamine-conjugated Donkey Anti-Rabbit IgG</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">711-025-152</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Cy 5-conjugated Donkey Anti-Guinea Pig IgG&#160;</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">706-175-148</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cy 5-conjugated Donkey Anti-Goat IgG</td>
			<td>Jackson ImmunoResearch</td>
			<td>705-175-147</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cy 5-conjugated Donkey Anti-Rabbit IgG</td>
			<td>Jackson ImmunoResearch</td>
			<td>711-175-152</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cy 5-conjugated Donkey Anti-Chicken IgG</td>
			<td>Jackson ImmunoResearch</td>
			<td style="width:147px;">703-175-155</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">DAPI</td>
			<td style="width:196px;">Millipore</td>
			<td style="width:147px;">S7113</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Disposable Reagent Reservoir 25 mL</td>
			<td style="width:196px;">Sorenson BioScience</td>
			<td style="width:147px;">39900</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">384 well, black/clear, tissue culture treated plate</td>
			<td style="width:196px;">BD Falcon</td>
			<td style="width:147px;">353962</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">96 well, black/clear, tissue culture treated plate</td>
			<td style="width:196px;">Costar</td>
			<td style="width:147px;">3603</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Multi-channel pipettor</td>
			<td style="width:196px;">Costar</td>
			<td style="width:147px;">4880</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">12-channel vaccume aspirator</td>
			<td style="width:196px;">Drummond</td>
			<td style="width:147px;">3-000-096</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Cell Scraper</td>
			<td style="width:196px;">Falcon</td>
			<td style="width:147px;">353085</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Isotemp Water Bath Model 2223&#160;</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:147px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">High-content screening instrument: ArrayScan VTI</td>
			<td style="width:196px;">Thermo Scientific</td>
			<td style="width:147px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

a high-content in vitro pancreatic islet &#946;-cell replication discovery platform

Loss of insulin-producing &#946;-cells is a central feature of diabetes. While a variety of potential replacement therapies are being explored, expansion of endogenous insulin-producing pancreatic islet &#946;-cells remains an attractive strategy. &#946;-cells have limited spontaneous regenerative activity; consequently, a crucial research effort is to develop a precise understanding of the molecular pathways that restrain &#946;-cell growth and to identify drugs capable of overcoming these restraints. Herein an automated high-content image-based primary-cell screening method to identify &#946;-cell replication-promoting small molecules is presented. Several, limitations of prior methodologies are surmounted. First, use of primary islet cells rather than an immortalized cell-line maximizes retention of in vivo growth restraints. Second, use of mixed-composition islet-cell cultures rather than a &#946;-cell-line allows identification of both lineage-restricted and general growth stimulators. Third, the technique makes practical the use of primary islets, a limiting resource, through use of a 384-well format. Fourth, detrimental experimental variability associated with erratic islet culture quality is overcome through optimization of isolation, dispersion, plating and culture parameters. Fifth, the difficulties of accurately and consistently measuring the low basal replication rate of islet endocrine-cells are surmounted with optimized immunostaining parameters, automated data acquisition and data analysis; automation simultaneously enhances throughput and limits experimenter bias. Notable limitations of this assay are the use of dispersed islet cultures which disrupts islet architecture, the use of rodent rather than human islets and the inherent limitations of throughput and cost associated with the use of primary cells. Importantly, the strategy is easily adapted for human islet replication studies. This assay is well suited for investigating the mitogenic effect of substances on &#946;-cells and the molecular mechanisms that regulate &#946;-cell growth.

For å vurdere β-celle eller α-celle-replikasjon, er en fire-fargers analyseprotokollen som kreves. Først blir gjenstander identifiseres ved DAPI farging (kanal 1, 386 nm). Deretter blir p-celler (event 1) telles: objekter som co-uttrykke PDX-1 + (kanal 2, 650 nm) og peri-atom insulin (kanal 3, 549 nm). Deretter blir replikere p-celler (hendelses 2) telles: P-celler (event 1) som co-express Ki-67 (kanal 4, 485 nm) (figur 3). Prosentandelen av replikerende p...

Watch this Scientific Journal Video about A High-content In Vitro Pancreatic Islet ÃŽÂ²-cell Replication Discovery Platform at JoVE.com

A High-content In Vitro Pancreatic Islet ÃƒÅ½Ã‚Â²-cell Replication Discovery Platform

Kritiske utfordringer for diabetes forskningsfeltet er å forstå de molekylære mekanismene som regulerer holmen β-celle replikering og å utvikle metoder for å stimulere β-celle gjenfødelse. Heri et høyt innhold av screening-metoden for å identifisere og vurdere β-celle-replikasjon-fremmende aktivitet av små molekyler er presentert.

En High-innhold<em&gt; In Vitro</em&gt; Bukspyttkjertelen Islet β-celle Replication Discovery Platform

Loss of insulin-producing &#946;-cells is a central feature of diabetes. While a variety of potential replacement therapies are being explored, expansion ...

Research

JoVE Journal

Medicine

A High-content In Vitro Pancreatic Islet &#946;-cell Replication Discovery Platform

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

High Content Screening in Neurodegenerative Diseases

Høyt innhold Screening i nevrodegenerative sykdommer

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Kirurgiske prosedyrer for en rotte modell av delvis Orthotopic levertransplantasjon med nedsatt arteriell Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Simulating Pancreatic Neuroplasticity: In Vitro Dual-neuron Plasticity Assay

Simulating Pancreatic Neuroplasticity: In Vitro Dual-neuron Plasticity Assay

Simulabukspyttkjertel Neuroplasticity:<em&gt; In Vitro</em&gt; Dual-nevron Plasticity analysen

Neuroplasticity is an inherent feature of the enteric nervous system and gastrointestinal (GI) innervation under pathological conditions. However, the ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Eksitotoksiske Stimulering av Hjerne Microslices som en<em&gt; In vitro</em&gt; Modell av Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

Modellering Spontan metastatisk nyrecellekarsinom (MRCC) hos mus etter Nephrectomy

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

Lepr<sup&gt; Db</sup&gt; Mus Modell av Type 2 Diabetes: Bukspyttkjertel Islet Isolasjon og Live-celle 2-Photon Imaging av intakte holmer

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, ...

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 ...

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

Laser Doppler: Et verktøy for måling bukspyttkjertelen Holme mikrovaskulær Vasomotion i Vivo

As a functional status of microcirculation, microvascular vasomotion is important for the delivery of oxygen and nutrients and the removal of carbon ...

Application of End-to-end Anastomosis in Robotic Central Pancreatectomy

Anvendelse av ende-til-ende anastomose i robot sentrale Pancreatectomy

Central pancreatectomy is carried out for the treatment of benign or low-malignant potential tumors located in the pancreatic neck or proximal part of ...