Name: in vitro colony assays for characterizing tri-potent progenitor cells isolated from the adult murine pancreas
Uploaded: 2016-06-10T15:00:00
Description: Watch this Scientific Journal Video about In Vitro Colony Assays for Characterizing Tri-potent Progenitor Cells Isolated from the Adult Murine Pancreas at JoVE.com

Vi takker Lucy Brown og Alexander Spalla fra Analytical cytometri Core på City of Hope for hjælp til sortering. Dette arbejde understøttes delvist af National Institutes of Health (NIH) giver R01DK081587 og R01DK099734 til HTK, og U01DK089533 til ADR, og af National Science Foundation tilskud NSF-DMR-1.206.121 og California Institute for regenerativ medicin tilskud RB5-07398 til DAT Understøtter fra Joseph J. Jacobs Institut for Molekylær Engineering for Medicin på Caltech til DAT, og dem fra Oxnard Foundation og Ella Fitzgerald Foundation til HTK er også taknemmeligt anerkendt. Finansiering: Dette arbejde understøttes delvist af National Institutes of Health (NIH) giver R01DK081587 og R01DK099734 til HTK, og U01DK089533 til ADR, og af National Science Foundation tilskud NSF-DMR-1.206.121 og California Institute for regenerativ medicin tilskud RB5-07398 til DAT Understøtter fra Joseph J. JacobsInstitut for Molekylær Engineering for Medicin på Caltech til DAT, og dem fra Oxnard Foundation og Ella Fitzgerald Foundation til HTK er også taknemmeligt anerkendt. Forskning rapporteret i denne publikation indgår arbejde udført i Analytisk cytometri Core og Light Microscopy Digital Imaging Core støttet af National Cancer Institute of National Institutes of Health under award nummer P30CA33572. Undersøgelse sponsor: Sponsor deltog ikke i undersøgelsen design, indsamling, analyse, eller fortolkningen af data.

Etisk Statement: Vi overholder de bredt accepterede etiske standarder i at udføre forskning for at sikre kvaliteten og integriteten af ​​resultaterne. Dyreforsøg er udført i overensstemmelse med protokoller, der er godkendt af Institutional Animal Care og brug Udvalg på City of Hope. 1. Forbered Single-celle Suspension fra Adult Murine pancreata BEMÆRK: I tidligere publikationer 7,11, blev CD-1 eller B6 baggrund mus anvendt; begge baggrunde gav lignende resultater. En transgene mus linj...

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De pancreas koloni assays og enkelt koloni analyser beskrevet her blev inspireret af de methylcellulose-holdige hæmatopoietisk koloni analyser, der har spillet store roller i decifrere biologi hæmatopoietiske stamceller i de seneste årtier 23. I disse assays (figur 5), dissocierede pancreasceller udplades i methylcellulose indeholdende halvfast medier med passende vækstfaktorer og ECM-proteiner, der understøtter dannelsen af Ring, tætte eller endokrine / Acinære kolonier 7. ...

Bugspytkirtlen er sammensat af tre store cellelinier; acinære celler udskiller fordøjelsesenzymer, kanaler udskiller mucin at afværge patogener og transportere fordøjelsesenzymer til tarmen, og endokrine celler udskiller hormoner, herunder insulin og glukagon, der opretholder glukose homøostase. Under fosterudviklingen i bugspytkirtlen, de tidlige duktale celler er kilden til tri-potente stamceller er i stand til at give anledning til de tre slægter i pancreas af voksne dyr 1,2. Fordi voksne stamceller og progenitorceller, såsom knoglemarvsstamceller, der allerede med succes anvendes til behandling af forskellige sygdomme 3, der er en intens interesse i at finde stilken, og progenitorceller i den voksne pancreas. Hvis isolering og manipulering af voksne pancreas stamceller og progenitorceller var muligt, kunne disse celler anvendes til at behandle sygdomme, såsom type 1 diabetes, hvor insulin-udskillende celler ødelægges af autoimmunitet. <p class = &quot;jove_content&quot;&gt; Uanset om tri-potente stamceller stadig eksisterer i voksne pancreas kanaler efter afslutningen af ​​fosterudviklingen er et spørgsmål, der er stærkt debatteret i det videnskabelige samfund. I denne debat og brug in vivo cre-lox afstamning-tracing teknikker, inada og kolleger viste, at voksne murine duktale celler mærket med en markør, kulsyreanhydrase II, kunne give anledning til alle tre pancreas afstamninger 4. Men ved hjælp andre duktale markører, såsom HNF1b 5 og Sox9 2, blev det konkluderet, at duktale celler ikke den største kilde til betaceller i voksne mus. For flere år siden, foreslog vi, at årsagen til den førnævnte debat kan skyldes manglen på området 6,7, af passende analytiske værktøjer, som kan bruges til at måle selvfornyelse og multi-slægt differentierings--to kriterier er nødvendige for at definere en stamcelle. In vivo cre-lox afstamning-tracing teknik nævntovenfor kan dokumentere for stamfader-afkom forhold på befolkningsniveau. Men denne slægt sporing teknik begrænset i sin magt for at skelne, om enkelte stamceller kan selv forny og differentiere i flere slægter. Single-celle analyse er vigtigt, fordi hvis flere mono-potente stamceller, hver med en forskellig afstamning potentiale, blev analyseret sammen, kan de tilsammen synes at have multi-slægt differentiering evner. Desuden stamceller er normalt en mindre population af en voksen organ. Aktiviteterne i en mindre celle population kunne være maskeret af den større population. Derfor er et negativt resultat fra en population undersøgelse ikke nødvendigvis viser forekomst af stamceller. Endelig er cre-lox afstamning sporing ikke tillader i øjeblikket måling af selv-fornyelse. For at begynde at behandle den tekniske kløft på området pancreasstamfadercelle biologi, koloni 7-11 eller organoide 12-15 </sup&gt; assays under anvendelse af 3D-kultur systemer blev udtænkt. To koloniassays for pancreas progenitorer blev udviklet i vores laboratorium: én indeholder et kommercielt præparat af murine ekstracellulære matrixproteiner (ECM) (se Metoder og udstyr tabel), og den anden indeholder laminin hydrogel, en defineret kunstig ECM protein 7-11. Progenitorceller blandes i halvfast medium indeholdende methylcellulose. Methylcellulose er et biologisk inert og tyktflydende materiale fremstillet af træfibre, og har været rutinemæssigt anvendt i hæmatopoietiske koloniassays 16. Den methylcellulose indeholdende halvfast medium begrænser bevægelsen af ​​enkelte progenitorceller, så de ikke kan re-aggregat. Men mediet er blød nok til at tillade en progenitorcelle at vokse og differentiere til en koloni af celler i 3D rummet. Efter traditionen af ​​hæmatologer, en pancreasstamfadercelle der var i stand til at give anledning til en koloni af celler var named en pancreas kolonidannende enhed (PCFU). PCFUs, når der dyrkes i murine ECM-holdige koloni assay, giver anledning til cystisk kolonier, der er opkaldt &quot;Ring&quot; kolonier 7. Ved tilsætning af et Wnt-agonist, R-spondin1, ind i den murine ECM-holdige kultur, nogle Ring kolonier bliver til &quot;Tætte&quot; kolonier 7. I denne artikel, er disse to typer af kolonier dyrket i murine ECM kultur samlet benævnt &quot;Ring / Tætte&quot; kolonier. Når Ring / Tætte kolonier dissocieret til enkelt celle suspension og re-belagte i kulturer, der indeholder laminin hydrogel, &quot;Endokrine / Acinære&quot; kolonier dannes 7. Brug enkelt koloni analyser, blev det konstateret, at størstedelen af Ring / Tætte og endokrine / Acinære kolonier, enten fra voksne (2-4 måneder gamle) 7,11 eller unge (1 uge gammel) 9 murine bugspytkirtel, udtrykker alle tre afstamning markører. Dette antyder, at de fleste af de PCFUs oprindelse er tri-potente. I det murine ECM-holdige koloni assay, voksne murine PCFUs håndfast selv forny og udvide cirka 500.000 gange over 11 uger i kultur 7. Murin ECM understøtter fortrinsvis differentieringen af duktale celler frem endokrine og acinære afstamninger, hvorimod i nærvær af laminin hydrogel, er murine PCFUs tilskyndes til at differentiere fortrinsvis i endokrine og acinære celler og i mindre grad til duktalt afstamning 7,9,11. Vigtigt er, Insulin + Glucagon - mono-hormonale celler genereres i laminin hydrogel kultur og udskille insulin i respons på glucose stimulation in vitro 7,9, hvilket tyder på funktionelle modenhed. Tri-slægt differentiering potentiale 7,9 og selv-fornyelse 11 af individuelle PCFUs bekræftes af encellede mikromanipulering, dvs. dyrkning én celle pr godt for kolonidannelse. Tilsammen udgør disse resultater beviser, at der er selvfornyende, tri-Potent, progenitor-lignende celler i postnatal murine bugspytkirtlen, der viser aktiviteterne i 3D kultur. De murine PCFU analyser beskrevet i denne artikel stammer fra en tidligere koloni assay designet til stamceller differentieret fra murine embryonale stamceller (mESCs) 17. Det protokol er dokumenteret i detaljer i en anden JOVE publikation 18. De kultur komponenter og teknikker, der kræves for at udføre den murine ECM-holdige koloni assay for voksne PCFUs er de samme som for Mesc-afledte stamfædre 17,18. Derfor vil disse aspekter af analysen ikke gentages her; i stedet følgende procedurer vil blive behandlet: 1) dissociation af den voksne bugspytkirtlen og sortering CD133 + Sox9 / EGFP + duktale celler, som beriger PCFUs fra voksne mus 7, 2) encellede manipulation af de sorterede celler, 3) single-koloni analyser ved hjælp microfluidic QRT-PCR og hel-mount immunfarvning, og 4) dissociation af Colonies mod enkelt-cellesuspension og re-plating i murine ECM eller laminin hydrogel koloniassays.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Murine ECM proteins (Matrigel)</td>
			<td>Becton Dickson (Franklin Lakes, NJ, USA)</td>
			<td>354230</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Laminin Hydrogel</td>
			<td>Provided by David Tirrell (Pasadena, CA USA)</td>
			<td></td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Methylcellulose</td>
			<td>Shinetsu Chemical (Tokyo, Japan)</td>
			<td></td>
			<td>1500 centipoise (dynamic viscosity unit equal to 15g/cm/s) (high viscosity)</td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s Phosphate-Buffered Saline</td>
			<td>Mediatech (Manassas, VA, USA)</td>
			<td>21-031-CV</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline</td>
			<td>Gibco (Grand Island, NY, USA)</td>
			<td>15070-063</td>
			<td></td>
		</tr>
		<tr>
			<td>50mL Flacon Conical vial</td>
			<td>Corning Inc. (Corning, NY, USA)</td>
			<td>352070</td>
			<td></td>
		</tr>
		<tr>
			<td>100mmx20mm Suspension culture dish</td>
			<td>Corning Inc. (Corning, NY, USA)</td>
			<td>430591</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td>A8412</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Gibco (Grand Island, NY, USA)</td>
			<td>15070-063</td>
			<td></td>
		</tr>
		<tr>
			<td>DNase1</td>
			<td>Calbiochem (Darmstadt, Germany)</td>
			<td>260913</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagenase B</td>
			<td>Roche (CH-4070, Basel, Schweiz, Switzerland)</td>
			<td>11088831001</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Anti-mouse CD16/32</td>
			<td>Biolegend (San Diego, CA, USA)</td>
			<td>101310</td>
			<td>low endotoxin, azide free</td>
		</tr>
		<tr>
			<td>PE-Cy7 Rat IgG2a &#954; Isotype Control</td>
			<td>Biolegend (San Diego, CA, USA)</td>
			<td>400522</td>
			<td></td>
		</tr>
		<tr>
			<td>Rat IgG1 &#954; Isotype Control</td>
			<td>eBioscience (San Diego, CA, USA)</td>
			<td>13-4301-82</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-CD133-Biotin</td>
			<td>eBioscience (San Diego, CA, USA)</td>
			<td>13-1331-82</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-CD71-PE-Cy7</td>
			<td>Biolegend (San Diego, CA, USA)</td>
			<td>113812</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptavidin-Allophycocyanin</td>
			<td>Biolegend (San Diego, CA, USA)</td>
			<td>405207</td>
			<td></td>
		</tr>
		<tr>
			<td>4&#39;,6-Diamidino-2-phenylindole</td>
			<td>Invitrogen (Waltham, MA,&#160; USA)</td>
			<td>3571</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Anti-mucin 1</td>
			<td>Thermo Fisher Scientific (Waltham, MA USA)</td>
			<td>HM-1630-P1</td>
			<td></td>
		</tr>
		<tr>
			<td>Dylight 649 Goat anti-Armenian Hamster</td>
			<td>Jackson Immuno (West Grove, PA, USA)</td>
			<td>127-495-160</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s Modified Eagle Medium: Nutrient Mixture F-12</td>
			<td>Mediatech(Manassas, VA, USA)</td>
			<td>10-092-CV</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Tissure Culture Biologicals (Long Beach, CA, USA)</td>
			<td>101</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Tris Ethylenediaminetetraacetic acid</td>
			<td>TEKnova (Hollister, CA, USA)</td>
			<td>T0221</td>
			<td></td>
		</tr>
		<tr>
			<td>Rneasy Micro Kit</td>
			<td>Qiagen (Venlo, Netherlands)</td>
			<td>74004</td>
			<td></td>
		</tr>
		<tr>
			<td>QuantiTec Reverse Transcription Kit</td>
			<td>Qiagen (Venlo, Netherlands)</td>
			<td>205310</td>
			<td></td>
		</tr>
		<tr>
			<td>CellsDirect One-Step qRT-PCR Kit</td>
			<td>Ambion/Invitrogen(Grand Island, NY, USA)</td>
			<td>11753-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Santa Cruz Bio (Santa Cruz, CA, USA)</td>
			<td>SC-281692</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat serum</td>
			<td>Jackson Immuno (West Grove, PA, USA)</td>
			<td>005-000-121</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Donkey serum</td>
			<td>Jackson Immuno (West Grove, PA, USA)</td>
			<td>0017-000-121</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td>T9284</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td>T-4799</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylenediaminetetraacetic acid</td>
			<td>Invitrogen (Waltham, MA,&#160; USA)</td>
			<td>15575-020</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin-EDTA</td>
			<td>Life Technologies (Waltham, MA, USA)</td>
			<td>25200-056</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Water</td>
			<td>Gibco (Grand Island, NY, USA)</td>
			<td>15230-147</td>
			<td>Molecular biology grade</td>
		</tr>
		<tr>
			<td>Pasteur Pipette</td>
			<td>Fisher Scientific (Pittsburgh, PA , USA)</td>
			<td>13-678-8B</td>
			<td></td>
		</tr>
		<tr>
			<td>40um Filter Mesh</td>
			<td>Fisher Scientific (Pittsburgh, PA , USA)</td>
			<td>08-771-1</td>
			<td></td>
		</tr>
		<tr>
			<td>70 um filter mesh</td>
			<td>Fisher Scientific (Pittsburgh, PA , USA)</td>
			<td>08-771-2</td>
			<td></td>
		</tr>
		<tr>
			<td>TC Plate 96 Well Suspension</td>
			<td>Sarstedt</td>
			<td>83.3924 (Previously 83.1835)</td>
			<td></td>
		</tr>
		<tr>
			<td>1cc Syringe</td>
			<td>Becton Dickson (Franklin Lakes, NJ, USA)</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr>
			<td>10cc Syringe</td>
			<td>Becton Dickson (Franklin Lakes, NJ, USA)</td>
			<td>301604</td>
			<td></td>
		</tr>
		<tr>
			<td>48.48 Dyanmic Array Chip</td>
			<td>Fluidigm (San Francisco, CA, USA)</td>
			<td>BMK-M-48.48</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluidigm GE 48.48 Dynamic Array Sample &#38; Assay Loading Reagent Kit</td>
			<td>Fluidigm (San Francisco, CA, USA)</td>
			<td>85000800</td>
			<td></td>
		</tr>
		<tr>
			<td>TaqMan Universal PCR Master Mix</td>
			<td>Applied Biosystems (Grand Island, NY, USA)</td>
			<td>4304437</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyethylene glycol sorbitan monolaurate</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td>P7949</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass Bottom Dish</td>
			<td>MatTek (Ashland, MA, USA)</td>
			<td>P35G-1.5-14-C</td>
			<td>35 mm petri dish with glass bottom</td>
		</tr>
		<tr>
			<td>Mouth Piece/ Rubber Tubing</td>
			<td>Renova Life Inc. (College Park, MD, USA)</td>
			<td>MP-SET</td>
			<td></td>
		</tr>
		<tr>
			<td>Nicotinamide</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td>N0636</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Vascular Endothelial Growth Factor</td>
			<td>R&#38;D Systems (Minneapolis, MN, USA)</td>
			<td>293-VE</td>
			<td>Stock kept at -80oC</td>
		</tr>
		<tr>
			<td>Activin B</td>
			<td>R&#38;D Systems (Minneapolis, MN, USA)</td>
			<td>659-AB</td>
			<td>Stock kept at -80oC</td>
		</tr>
		<tr>
			<td>Extendin 4</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td>E7144</td>
			<td>Stock kept at -20oC</td>
		</tr>
		<tr>
			<td>Rspondin-1</td>
			<td>R&#38;D Systems (Minneapolis, MN, USA)</td>
			<td>3474-RS</td>
			<td>Stock kept at -80oC</td>
		</tr>
		<tr>
			<td>Falcon 5mL Polystyrene Round-Bottom Tube</td>
			<td>Corning Inc. (Corning, NY, USA)</td>
			<td>352054</td>
			<td></td>
		</tr>
		<tr>
			<td>PrecisionGlide Needle 18Gx1 1/2</td>
			<td>Becton Dickson (Franklin Lakes, NJ, USA)</td>
			<td>305196</td>
			<td></td>
		</tr>
		<tr>
			<td>PrecisionGlide Needle 16Gx1 1/2</td>
			<td>Becton Dickson (Franklin Lakes, NJ, USA)</td>
			<td>305198</td>
			<td></td>
		</tr>
		<tr>
			<td>Costar Ultra-Low Attachment Surface 24 well flat bottom plate&#160;</td>
			<td>Corning Inc. (Corning, NY, USA)</td>
			<td>3473</td>
			<td></td>
		</tr>
		<tr>
			<td>Costar 96 Black Well Plate</td>
			<td>Corning Inc. (Corning, NY, USA)</td>
			<td>3603</td>
			<td>Flat, clear bottom with lid.&#160; Black polystyrene TC-treated microplates</td>
		</tr>
		<tr>
			<td>Zeiss LSM510 META NLO Axiovert 200M Inverted Microscope</td>
			<td>Carl Zeiss AG (Oberkochen, Germany)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Biomark HD&#160;</td>
			<td>Fluidigm (San Francisco, CA, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Aria Special Order Research Product Cell Sorter</td>
			<td>Becton Dickson (Franklin Lakes, NJ, USA)</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

in vitro colony assays for characterizing tri-potent progenitor cells isolated from the adult murine pancreas

Stem og stamceller fra voksne bugspytkirtlen kunne være en potentiel kilde til terapeutiske beta-lignende celler til behandling af patienter med type 1-diabetes. Det er dog stadig uvist, om der findes stamceller og progenitorceller i den voksne pancreas. Forskning strategier via cre-lox afstamning-sporing i voksne mus har givet resultater, som enten støtte eller afkræfte den idé, at betaceller kan genereres fra kanalerne, den formodede sted, hvor voksne bugspytkirtlen stamfædre kan opholde. Disse in vivo cre-lox slægt-tracing metoder kan imidlertid ikke besvare spørgsmål fra selvfornyelse og multi-slægt differentiering-to kriterier, der er nødvendige for at definere en stamcelle. Til at begynde at behandle dette tekniske hul, vi udtænkt 3-dimensionelle koloni assays for bugspytkirtlen forfædre. Kort efter vores indledende offentliggørelse, andre laboratorier uafhængigt udviklet en lignende, men ikke identiske, metode kaldet organoide assay. Sammenlignet med den organoide assay vores fremgangsmåde anvendermethylcellulose, der danner viskose opløsninger, som tillader optagelse af ekstracellulære matrixproteiner ved lave koncentrationer. De methylcellulose-holdige analyser tillader lettere påvisning og analyser af stamceller på enkelt-celle niveau, som er afgørende, når stamfædre udgør en lille sub-population, som det er tilfældet for mange voksne orgel stamceller. Sammen resultater fra flere laboratorier demonstrerer in vitro selvfornyelse og multi-slægt differentiering af pancreasstamfaderceller-lignende celler fra mus. De nuværende protokoller beskriver to methylcellulose-baserede koloniassays at karakterisere muse pancreas progenitorer; Den ene indeholder et kommercielt præparat af murine ekstracellulære matrixproteiner, og den anden en kunstig ekstracellulært matrixprotein kendt som en laminin hydrogel. De her viste teknikker er 1) dissociation af bugspytkirtlen og sortering af CD133 + Sox9 / EGFP + duktale celler fra voksne mus, 2) enkelt celle manipulation af sorted celler, 3) enkelt koloni analyser bruger mikrofluid QRT-PCR og hel-mount immunfarvning, og 4) dissociation af primære kolonier i encellede suspensioner og re-plating til sekundære koloni analyser til at vurdere selvfornyelse eller differentiering.

Voksne pancreasstamfaderceller kan beriges ved fluorescens-aktiveret cellesortering (figur 1). Den Sox9 / EGFP transgene mus linje bruges her blev først frembringes som resultat af GENSAT Brain Atlas Project 19 og den EGFP reporter er under kontrol af en bakteriel kunstigt kromosom indeholdende ~ 75 kb opstrøms og ~ 150 kb nedstrøms sekvenser af Sox9 20 . I disse mus, EGFP etiketter pancreas kanaler effektivt og specifikt 22. Pancreas ...

Watch this Scientific Journal Video about In Vitro Colony Assays for Characterizing Tri-potent Progenitor Cells Isolated from the Adult Murine Pancreas at JoVE.com

In Vitro Colony Assays for Characterizing Tri-potent Progenitor Cells Isolated from the Adult Murine Pancreas

In vitro koloni assays til påvisning selvfornyelse og differentiering af stamceller isoleret fra voksne murine bugspytkirtlen er udformet. I disse assays pancreatiske progenitorer medfører cellekolonier i 3-dimensionalt rum i methylcellulose indeholdende halvfast medium. Protokoller for håndtering enkelte celler og karakterisering af de enkelte kolonier beskrives.

In vitro Colony Analyser til karakterisering Tri-potente progenitorceller isoleret fra voksne murine Pancreas

Stem and progenitor cells from the adult pancreas could be a potential source of therapeutic beta-like cells for treating patients with type 1 diabetes. ...

Research

JoVE Journal

Developmental Biology

In Vitro Colony Assays for Characterizing Tri-potent Progenitor Cells Isolated from the Adult Murine Pancreas

Derivation of Cardiac Progenitor Cells from Embryonic Stem Cells

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Production and Use of Lentivirus to Selectively Transduce Primary Oligodendrocyte Precursor Cells for In Vitro Myelination Assays

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Adult rat and human spinal cord neural stem/progenitor cells (NSPCs) cultured in growth factor-enriched medium allows for the proliferation of ...

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Characterizing thymic settling progenitors is important to understand the pre-thymic stages of T cell development, essential to devise strategies for T ...

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This manuscript describes a means to enrich for neural progenitors from the marrow stromal cell (MSC) population and thereafter to direct them to the ...

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Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the ...

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