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 protokol blev udført i overensstemmelse med Institutional Animal Care og brug Udvalg (IACUC) fra Stanford University School of Medicine. Den beskrevne protokol er skaleret til holm isolation fra seks 250 - 300 g (8 - 9 uger gamle) Sprague Dawley-rotter, som er tilstrækkelig til at frembringe 228 brønde i en plade med 384 brønde for islet-cellereplikation vurdering. 1. Klargøring <ol><li> Forbered coating medier før initiering holm isolering ved opsamling af konditionerede medier af 804G ro...

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Eksperimentelle metoder til at studere de molekylære veje, der styrer β-celle vækst og regeneration er vigtige redskaber for diabetes forskere. Heri, at en rotte-ø-baserede screening platform identificere og karakterisere små molekyler stimulatorer af β-celle replikation præsenteres. Mens de fleste aspekter af denne protokol nemt udføres af erfarne forskere, et par skridt kræver særlig teknik. Først under islet isolation, kanylering af galde-kanal uden at forstyrre dens integritet...

Diabetes omfatter en samling af lidelser, der deler den fælles endepunkt af forstyrret glukose homøostase. Selvom de patogene mekanismer diabetes undertyper er forskellige, de deler konsekvensen af nedsat β-cellemasse, dvs. tab af insulin produktionskapacitet 1,2. I øjeblikket, diabetes behandlingsstrategier stole på kronisk tilføre insulin, farmakologiske stimulering af insulinproduktion eller forbedring af insulinfølsomheden, og sjældent, at transplantation af pancreas-øer eller hel bugspytkirtlen 3,4. Desværre succes af disse strategier er kortvarig og / eller ikke i tilstrækkelig grad rekapitulere funktionen af ​​endogene insulinproduktion. Trods nytten af ​​at udvikle en metode til at stimulere β-celle regenerering, findes der ingen sådan fremgangsmåde. Derfor er en større diabetesforskning mål er at udvikle metoder til at generere nye p-celler eller til at udvide endogen β-cellemasse 5 </sup&gt;. Selvom β-celle regenerering fra vedvarende energikilder såsom fosterstamceller skrider, sikkerhed og effektivitet bekymringer gør udøvelse af alternative strategier, herunder udvidelse af modne p-celler, en prioritet 6,7. Vigtigt er det, den dominerende kilde til nye p-celler in vivo er allerede eksisterende p-celler frem for specialiserede stamceller 8,9. Selvom p-celler synes at have begrænset replikation kapacitet, en lille stigning i β-cellemasse (~ 30%) kan være tilstrækkelig til at genoprette glucosehomøostase i mange diabetikere. Endvidere in situ farmakologiske stimulering af β-cellemassen er en potentielt billig og skalerbar behandlingsstrategi. Heri et højt indhold screeningsfremgangsmåde til identifikation og karakterisering små molekyler, der stimulerer β-cellevækst præsenteres. En række forskellige in vitro eksperimentelle fremgangsmåder kan anvendes til at identificere genprodukter og / eller molekyler that fremme primær β-cellereplikation. Tidlige indsats til måling β-celle replikation induktion brugt føtal gnaver pancreas kultur eller intakte isoleret ø kulturer til at måle [3H] thymidin, BrdU inkorporering eller mitotiske organer i aldehyd-thionin eller insulin farvede befolkning som reaktion på specifikke behandlingsmetoder vilkår 10, 11. Disse in vitro-metoder og varianter heraf har flere begrænsninger. Fremtrædende mangler omfatter (1) anvendelse af føtale celler, som, i modsætning til modne p-celler, udviser en høj basal β-celle replikation sats og er vækst reguleres i en særskilt måde 12; (2) den subjektive karakter af forsøgslederen-afhængige pådømmelse af β-celle replikation begivenheder; (3) arbejds- og tidskrævende karakter af forsøgslederen-afhængige optælling af β-celle replikation begivenheder forsinker eksperimentel gennemløb; (4) brugen af ​​nuklear iblanding / plet / udseende for at identificere replikation selvts og en ikke-overlappende cytoplasmatisk farvning til identifikation p-celler fører til misattribution af nærliggende ikke-β-celle replikeringsaktiviteter til p-celler. For nylig modne primære P-celler er blevet anvendt til at vurdere konsekvenserne af transgen overekspression samt gen-produkt eller sammensatte behandling på β-celle replikation 13-16. Imidlertid har disse undersøgelser også påberåbes subjektive optælling af replikation begivenheder, cytoplasmatisk staining- eller ikke-specifikke metoder til β-celle identifikation og / eller arbejdsintensive trin, der begrænser gennemløb, fx individuel slide-brønd plating af celler eller intakte ø paraffinindlejring og forarbejdning 17. Især en billedbaseret human β-cellereplikation screening metode, der ligner den heri præsenterede, er blevet offentliggjort 18; imidlertid vellykkede anvendelse af dette assay ikke påvist, og anvendelsen af ​​humane øer til primær screening kan ikke være stort set fealigt. En alternativ strategi til identificering replikations-fremmende stoffer er at vurdere væksten induktion af β-cellelinier. Indledende indsats brugt transformerede Ø-celle-linjer såsom min6 celler eller INS 832/13-celler 14,19-21. Men disse cellelinjer demonstrere uhæmmet vækst, og har megen lighed med godt differentierede P-celler 22. Derfor vækst-induktion kapacitet er minimal, af uklare relevans og til tider vanskeligt at rekapitulere. En forbedret strategi for celle-line baseret screening udnytter &quot;reversibelt forvandlet&quot; celler, der er vækst anholdt i fravær af tetracyclin (doxycyclin) -afhængig SV40 T-antigen udtryk 23,24. Det er imidlertid uklart, om disse celler tilbage til en &quot;normal&quot; β-celle-lignende tilstand efter doxycyclin fjernelse. Uheldigvis har anvendelse af disse celler gav generaliserede vækstfremmende forbindelser, der ikke synes at have umiddelbar anvendelighed24. Samlet set brugen af cellelinier til at undersøge vækstregulering af en celletype udviser minimal spontan replikationsaktivitet kan have begrænset anvendelighed. Den β-cellereplikation screening platform præsenteret heri benytter modne primære rotte P-celler til at bibeholde i reguleringen vivo væksten i videst muligt omfang, islet-cellekulturer af sammensætningen blandet celletype at muliggøre identifikation af slægt-begrænset vækstfremmende aktiviteter, multi brønds formatering at maksimere gennemløb og automatiseret analyse at eliminere bias og lette gennemløb. Vellykket anvendelse af denne platform har muliggjort identifikation af adskillige forbindelser, der fremmer β-cellereplikation 25,26. Derudover er assayet blevet anvendt til struktur-aktivitets-relationer undersøgelser og kemiske epistasis eksperimenter for at tilvejebringe mekanistiske indsigt i den molekylære regulering af β-cellereplikation. Den præsenterede platform lykkedes tilpasset for lentivirale RNAi-baserede undersøgelse af β-celle replikation veje 25. Begrænsninger af assayet indbefatter begrænset skalerbarhed (anvendelse af primære celler), anvendelse af gnaver snarere end humane ø-celler (selvom assayet kan tilpasses for humane ø-undersøgelser), udgifter forbundet med antistof-baserede billeddannelse og primær holm brug, anvendelse af spredte holme (forstyrret holm arkitektur) for at lette automatiseret erhvervelse image og afhængighed af tilgængeligheden af ​​en automatiseret mikroskop med erhvervelse image og analysekapacitet. Selv om en let in vivo screening metode til identificering af genprodukter eller forbindelser, som stimulerer β-celle regenerering in situ ville være ideelt, sådan platform er endnu ikke tilgængelig 27. Følgelig den beskrevne platform er egnet til forsker interesseret i at undersøge de fleste aspekter af β-cellereplikation.

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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>
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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>
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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>
		<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 at vurdere β-celle eller α-celle replikation, er en fire-farve assay-protokol kræves. For det første er objekter identificeret ved DAPI farvning (kanal 1, 386 nm). Dernæst p-celler (event 1) tælles: genstande, der co-udtrykker PDX-1 + (kanal 2, 650 nm) og peri-nuklear insulin (kanal 3, 549 nm). Efterfølgende replikerende p-celler (event 2) tælles: P-celler (event 1) at co-udtrykker Ki-67 (kanal 4, 485 nm) (figur 3). Procentdelen af ​​replikeren...

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 udfordringer for diabetes forskningsfelt er at forstå de molekylære mekanismer, der regulerer holm β-celle replikation og at udvikle metoder til at stimulere β-celle regenerering. Heri et højt indhold screeningsmetode til at identificere og vurdere β-celle replikation-fremmende aktivitet af små molekyler er præsenteret.

En High-indhold<em&gt; In vitro</em&gt; Pancreas 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

Højt indhold Screening i neurodegenerative sygdomme

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 procedurer for en rottemodel af partiel ortotopisk levertransplantation med hepatisk arteriel Genopbygning

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

Simulering Pancreas Neuroplasticitet:<em&gt; In Vitro</em&gt; Dual-neuron Plasticitet Assay

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

Excitotoksiske Stimulering af Brain Microslices som en<em&gt; In vitro</em&gt; Model of 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

Modeling Spontan metastaserende renalcellecarcinom (MRCC) i mus efter nefrektomi

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 Model af type 2 diabetes: Pancreas Islet Isolering og live-cell 2-Photon Imaging af intakte Islets

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

Studere i bugspytkirtlen Kræft stamceller Karakteristik for udvikling af 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 værktøj til måling af pancreas Islet mikrovaskulære hjerte In 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 af ende-til-ende anastomose i robot centrale 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 ...