Name: Isolatie en zuivering van het pancreaseiland door Lewisratten met behulp van enzymatische vertering en dichtheidsgradiëntscheiding (Video) | JoVE Journal
Uploaded: 2026-04-03T12:53:00.000+00:00
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Description: Leer een gestandaardiseerd protocol voor de isolatie en zuivering van pancreaseilanden uit Lewis-ratten met behulp van enzymatische vertering en dichtheidsgradiëntscheiding. Deze methode optimaliseert opbrengst, zuiverheid en levensvatbaarheid voor diabetesonderzoek.

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<ol><li>Grattoni, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Harnessing+cellular+therapeutics+for+type+1+diabetes+mellitus%3A+progress%2C+challenges%2C+and+the+road+ahead%5BTitle%5D)+AND+%22Nat+Rev+Endocrinol%22%5BJournal%5D)">Harnessing cellular therapeutics for type 1 diabetes mellitus: progress, challenges, and the road ahead.</a> Nat Rev Endocrinol. 21 (1), 14-30 (2025).</li>
<li>Páez-Mayorga, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Emerging+strategies+for+%CE%B2+cell+transplantation+to+treat+type+1+diabetes+mellitus%5BTitle%5D)+AND+%22Cell+Transplant%22%5BJournal%5D)">Emerging strategies for β cell transplantation to treat type 1 diabetes mellitus.</a> Cell Transplant. 31, 963689721004(2022).</li>
<li>Cabrera, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+unique+cytoarchitecture+of+human+pancreatic+islets+and+its+relevance+to+%CE%B2-cell+function%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">The unique cytoarchitecture of human pancreatic islets and its relevance to β-cell function.</a> Proc Natl Acad Sci U S A. 103 (7), 2334-2339 (2006).</li>
<li>Brissova, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pancreatic+islet+microvasculature+and+blood+flow%5BTitle%5D)+AND+%22J+Clin+Invest%22%5BJournal%5D)">Pancreatic islet microvasculature and blood flow.</a> J Clin Invest. 124 (4), 1871-1883 (2014).</li>
<li>Jiang, F. X., Morahan, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pancreatic+islet+architecture+and+function%5BTitle%5D)+AND+%22Cell+Death+Discov%22%5BJournal%5D)">Pancreatic islet architecture and function.</a> Cell Death Discov. 10, 170(2024).</li>
<li>Sato, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hypoxia+induces+%CE%B2-cell+apoptosis+and+functional+suppression+in+isolated+islets%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Hypoxia induces β-cell apoptosis and functional suppression in isolated islets.</a> PLoS One. 9 (12), e114868(2014).</li>
<li>Tiedge, M., Lortz, S., Drinkgern, J., Lenzen, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Relation+between+antioxidant+enzyme+gene+expression+and+antioxidative+defense+status+of+insulin-producing+cells%5BTitle%5D)+AND+%22Diabetes%22%5BJournal%5D)">Relation between antioxidant enzyme gene expression and antioxidative defense status of insulin-producing cells.</a> Diabetes. 46 (11), 1733-1742 (1997).</li>
<li>Ichii, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Shipment+of+human+islets+for+transplantation%5BTitle%5D)+AND+%22Am+J+Transplant%22%5BJournal%5D)">Shipment of human islets for transplantation.</a> Am J Transplant. 7 (4), 1010-1020 (2007).</li>
<li>Carobbio, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rapid+GSIS+kinetics+in+freshly+isolated+mouse+islets%5BTitle%5D)+AND+%22Diabetologia%22%5BJournal%5D)">Rapid GSIS kinetics in freshly isolated mouse islets.</a> Diabetologia. 47 (9), 1652-1662 (2004).</li>
<li>Zawalich, W. S., Zawalich, K. C., Kelley, G. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Immediate+perifusion+of+fresh+rat+and+mouse+islets%5BTitle%5D)+AND+%22Am+J+Physiol+Endocrinol+Metab%22%5BJournal%5D)">Immediate perifusion of fresh rat and mouse islets.</a> Am J Physiol Endocrinol Metab. 294 (2), E444-E450 (2008).</li>
<li>Wassmer, C. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Clinical+use+of+freshly+isolated+islets+transplanted+within+48+h%5BTitle%5D)+AND+%22Transplant+Proc%22%5BJournal%5D)">Clinical use of freshly isolated islets transplanted within 48 h.</a> Transplant Proc. 53 (6), 1907-1912 (2021).</li>
<li>Tai, D. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Immediate+autotransplantation+of+islets+following+total+pancreatectomy%5BTitle%5D)+AND+%22JAMA+Surg%22%5BJournal%5D)">Immediate autotransplantation of islets following total pancreatectomy.</a> JAMA Surg. 150 (5), 472-479 (2015).</li>
<li>Carter, J. D., Dula, S. B., Corbin, K. L., Wu, R., Nunemaker, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+practical+guide+to+rodent+islet+isolation+and+assessment%5BTitle%5D)+AND+%22Biol+Proced+Online%22%5BJournal%5D)">A practical guide to rodent islet isolation and assessment.</a> Biol Proced Online. 11, 3-31 (2009).</li>
<li>Emamaullee, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lewis+rats+in+transplantation+studies%5BTitle%5D)+AND+%22Diabetes%22%5BJournal%5D)">Lewis rats in transplantation studies.</a> Diabetes. 65 (5), 1350-1359 (2016).</li>
<li>de Groot, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rat+islet+isolation+yield+and+function+are+donor+strain+dependent%5BTitle%5D)+AND+%22Lab+Anim%22%5BJournal%5D)">Rat islet isolation yield and function are donor strain dependent.</a> Lab Anim. 38 (2), 200-206 (2004).</li>
<li>Moazenchi, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Strain+variability+in+islet+yield+and+isolation%5BTitle%5D)+AND+%22Islets%22%5BJournal%5D)">Strain variability in islet yield and isolation.</a> Islets. 2 (5), 283-289 (2010).</li>
<li>Dolenšek, J., Slak Rupnik, M., Stožer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation+variability+limits+reproducibility+in+rat+islet+studies%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Isolation variability limits reproducibility in rat islet studies.</a> PLoS One. 14 (5), e0216136(2019).</li>
<li>Stehno-Bittel, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+flaws+and+future+of+islet+volume+measurements%5BTitle%5D)+AND+%22Cell+Transplant%22%5BJournal%5D)">The flaws and future of islet volume measurements.</a> Cell Transplant. 27 (4), 571-578 (2018).</li>
<li>Kissler, H. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Validation+of+methodologies+for+quantifying+isolated+human+islets%5BTitle%5D)+AND+%22Cell+Transplant%22%5BJournal%5D)">Validation of methodologies for quantifying isolated human islets.</a> Cell Transplant. 18 (6), 611-620 (2009).</li>
<li>Papas, K. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Islet+assessment+for+transplantation%5BTitle%5D)+AND+%22Cell+Transplant%22%5BJournal%5D)">Islet assessment for transplantation.</a> Cell Transplant. 18 (6), 519-528 (2009).</li>
<li>Dono, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Low+temperature+collagenase+digestion+for+islet+isolation+from+48-hour+cold-preserved+rat+pancreas%5BTitle%5D)+AND+%22Transplantation%22%5BJournal%5D)">Low temperature collagenase digestion for islet isolation from 48-hour cold-preserved rat pancreas.</a> Transplantation. 57 (1), 22-26 (1994).</li>
<li>Shapiro, A. M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High+yield+of+rodent+islets+with+intraductal+collagenase+distension+and+warm+incubation%5BTitle%5D)+AND+%22Cell+Transplant%22%5BJournal%5D)">High yield of rodent islets with intraductal collagenase distension and warm incubation.</a> Cell Transplant. 5 (6), 631-638 (1996).</li>
<li>Corbin, K. L., Hughes, J. W., Nunemaker, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+practical+guide+to+rodent+islet+isolation+and+assessment+revisited%5BTitle%5D)+AND+%22Biol+Proced+Online%22%5BJournal%5D)">A practical guide to rodent islet isolation and assessment revisited.</a> Biol Proced Online. 23, 10(2021).</li>
<li>de Haan, B. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Factors+influencing+isolation+of+functional+pancreatic+rat+islets%5BTitle%5D)+AND+%22Pancreas%22%5BJournal%5D)">Factors influencing isolation of functional pancreatic rat islets.</a> Pancreas. 29 (1), e15-e22 (2004).</li>
<li>Kenyon, N. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Long-term+survival+and+function+of+intrahepatic+islet+allografts+in+baboons+treated+with+humanized+anti-CD154%5BTitle%5D)+AND+%22Diabetes%22%5BJournal%5D)">Long-term survival and function of intrahepatic islet allografts in baboons treated with humanized anti-CD154.</a> Diabetes. 48 (7), 1473-1481 (1999).</li>
<li>Kenyon, N. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Long-term+survival+and+function+of+intrahepatic+islet+allografts+in+rhesus+monkeys+treated+with+humanized+anti-CD154%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Long-term survival and function of intrahepatic islet allografts in rhesus monkeys treated with humanized anti-CD154.</a> Proc Natl Acad Sci U S A. 96 (14), 8132-8137 (1999).</li>
<li>Liberase T-Flex Research Grade - Method Sheet. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=%22eLabDoc%22">eLabDoc</a>. , Roche Diagnostics GmbH. (2025).</li>
<li>Miyazaki, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Strategy+towards+tailored+donor+tissue-specific+pancreatic+islet+isolation%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Strategy towards tailored donor tissue-specific pancreatic islet isolation.</a> PLoS One. 14 (5), e0216136(2019).</li>
<li>Cross, S. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Key+matrix+proteins+within+the+pancreatic+islet+basement+membrane+are+differentially+digested+during+human+islet+isolation%5BTitle%5D)+AND+%22Am+J+Transplant%22%5BJournal%5D)">Key matrix proteins within the pancreatic islet basement membrane are differentially digested during human islet isolation.</a> Am J Transplant. 17 (2), 451-461 (2017).</li>
<li>O'Gorman, D., Kin, T., Pawlick, R., Imes, S., Senior, P. A., Shapiro, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Clinical+islet+isolation+outcomes+with+a+highly+purified+neutral+protease+for+pancreas+dissociation%5BTitle%5D)+AND+%22Islets%22%5BJournal%5D)">Clinical islet isolation outcomes with a highly purified neutral protease for pancreas dissociation.</a> Islets. 5 (3), 111-115 (2013).</li>
<li>Wang, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adjustment+of+digestion+enzyme+composition+improves+islet+isolation+outcome+from+marginal+grade+human+donor+pancreata%5BTitle%5D)+AND+%22Cell+Tissue+Bank%22%5BJournal%5D)">Adjustment of digestion enzyme composition improves islet isolation outcome from marginal grade human donor pancreata.</a> Cell Tissue Bank. 8, 187-194 (2007).</li>
<li>McCall, M. D., Maciver, A. H., Pawlick, R., Edgar, R., Shapiro, A. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Histopaque+provides+optimal+mouse+islet+purification+kinetics%3A+comparison+study+with+Ficoll%2C+iodixanol+and+dextran%5BTitle%5D)+AND+%22Islets%22%5BJournal%5D)">Histopaque provides optimal mouse islet purification kinetics: comparison study with Ficoll, iodixanol and dextran.</a> Islets. 3 (4), 144-149 (2011).</li>
<li>Ricordi, C., Lacy, P. E., Finke, E. H., Olack, B. J., Scharp, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Automated+method+for+isolation+of+human+pancreatic+islets%5BTitle%5D)+AND+%22Diabetes%22%5BJournal%5D)">Automated method for isolation of human pancreatic islets.</a> Diabetes. 37 (4), 413-420 (1988).</li>
<li>BenchChem Technical Support Team. An in-depth technical guide to cell separation using iodixanol. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=%22BenchChem%22">BenchChem</a>. , (2026).</li>
<li>Dellê, H., Saito, M. H., Yoshimoto, P. M., Noronha, I. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+use+of+iodixanol+for+the+purification+of+rat+pancreatic+islets%5BTitle%5D)+AND+%22Transplant+Proc%22%5BJournal%5D)">The use of iodixanol for the purification of rat pancreatic islets.</a> Transplant Proc. 39 (2), 467-469 (2007).</li>
<li>Noguchi, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Iodixanol-controlled+density+gradient+during+islet+purification+improves+recovery+rate+in+human+islet+isolation%5BTitle%5D)+AND+%22Transplantation%22%5BJournal%5D)">Iodixanol-controlled density gradient during islet purification improves recovery rate in human islet isolation.</a> Transplantation. 87 (11), 1629-1635 (2009).</li>
<li>Min, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Superiority+of+Visipaque+%28iodixanol%29-controlled+density+gradient+over+Ficoll-400+in+adult+porcine+islet+purification%5BTitle%5D)+AND+%22Transplant+Proc%22%5BJournal%5D)">Superiority of Visipaque (iodixanol)-controlled density gradient over Ficoll-400 in adult porcine islet purification.</a> Transplant Proc. 42 (5), 1825-1829 (2010).</li>
<li>Paez-Mayorga, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neovascularized+implantable+cell+homing+encapsulation+platform+with+tunable+local+immunosuppressant+delivery+for+allogeneic+cell+transplantation%5BTitle%5D)+AND+%22Biomaterials%22%5BJournal%5D)">Neovascularized implantable cell homing encapsulation platform with tunable local immunosuppressant delivery for allogeneic cell transplantation.</a> Biomaterials. 257, 120232(2020).</li>
<li>Paez-Mayorga, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Implantable+niche+with+local+immunosuppression+for+islet+allotransplantation+achieves+type+1+diabetes+reversal+in+rats%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats.</a> Nat Commun. 13, 7951(2022).</li>
<li>Piemonti, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Islet+transplantation%5BTitle%5D)+AND+%22Endotext%22%5BJournal%5D)">Islet transplantation.</a> Endotext. , (2022).</li>
<li>Patel, S. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+foundation+for+engineering+a+pancreatic+islet+niche%5BTitle%5D)+AND+%22Front+Endocrinol%22%5BJournal%5D)">The foundation for engineering a pancreatic islet niche.</a> Front Endocrinol. 13, 881525(2022).</li>
</ol>

<table><tbody><tr><td>21 G x 2 in. BD PrecisionGlide Needle</td><td>BD</td><td>305129</td><td></td></tr><tr><td>500 &micro;m Brass Frame and 304 Stainless Steel Screen, 3" Diameter x 1" Deep (Sieve number 35)</td><td>McMaster-Carr</td><td>34735K525</td><td></td></tr><tr><td>Air-Tite Sterile Syringes, Luer lock</td><td>Fisher Scientific</td><td>14-817-178</td><td></td></tr><tr><td>Biogel PI UltraTouch M</td><td>M&ouml;lnlycke</td><td>42665</td><td></td></tr><tr><td>Bonn Scissors; Straight; Sharp Points; 15 mm Blade Length; 3 1/2" Overall Length</td><td>Roboz</td><td>RS-5840</td><td></td></tr><tr><td>Braintree Scientific GERMINATOR 500 DRY STERIL</td><td>Fisher Scientific</td><td>NC9956482</td><td></td></tr><tr><td>Cell-IQ Series 5.8 cu.ft. CO2 Laboratory Incubator</td><td>PHCBI</td><td>MCO-170AICUVHL-PA</td><td></td></tr><tr><td>Corning 100 mm Not TC-treated Culture Dish</td><td>Corning</td><td>430591</td><td></td></tr><tr><td>Corning Fetal Bovine Serum, 500 mL, Regular, USDA Safety Tested (Heat Inactivated)</td><td>Corning</td><td>35011CV</td><td></td></tr><tr><td>Corning Islet Gradient 1.069 (density 1.069 g/cm3)</td><td>Corning</td><td>99-815-CIS</td><td>Vereist special verzoek van bedrijf</td></tr><tr><td>Corning Islet Gradient 1.096 (density 1.096 g/cm3)</td><td>Corning</td><td>99-691-CIS</td><td>Vereist special verzoek van bedrijf</td></tr><tr><td>Corning Islet Gradient 1.108 (density 1.108 g/cm3)</td><td>Corning</td><td>99-692-CIS</td><td>Vereist special verzoek van bedrijf</td></tr><tr><td>Covidien Tendersorb Underpads</td><td>Fisher Scientific</td><td>22-225-102</td><td></td></tr><tr><td>DNase I Grade II</td><td>Roche</td><td>10104159001</td><td></td></tr><tr><td>Falcon&nbsp;50 mL High Clarity Conical Centrifuge Tubes</td><td>Fisher Scientific</td><td>14-432-22</td><td></td></tr><tr><td>Fisherbrand Aluminum Foil, Standard-Gauge Roll</td><td>Fisher Scientific</td><td>01-213-100</td><td></td></tr><tr><td>Fisherbrand&nbsp;Nonwoven Gauze Sponges</td><td>Fisher Scientific</td><td>22-028-559</td><td></td></tr><tr><td>Fisherbrand Reusable Glass Heavy-Duty Low-Form Beakers</td><td>Fisher Scientific</td><td>FB101600</td><td></td></tr><tr><td>Fluorescence Mircoscope&nbsp; BZ-X810</td><td>Keyence</td><td>BZ-X810</td><td></td></tr><tr><td>Glucose Solution</td><td>Gibco</td><td>A2494001</td><td></td></tr><tr><td>Halsey Micro Needle Holder</td><td>Fine Science Tools</td><td>12500-12</td><td></td></tr><tr><td>HBSS, calcium, magnesium, no phenol red</td><td>Gibco</td><td>14025092</td><td></td></tr><tr><td>HEPES (1 M)</td><td>Gibco</td><td>15630080</td><td></td></tr><tr><td>Hemostatic Forceps Classic Crile 5-1/2 Inch Length</td><td>McKesson</td><td>1231217</td><td></td></tr><tr><td>Hudson (Ewald) Forceps</td><td>Roboz</td><td>RS-5238</td><td></td></tr><tr><td>Hypodermic Needle Monoject 1-1/2 Inch Length 16 Gauge Regular Wall Without Safety</td><td>McKesson</td><td>43481</td><td></td></tr><tr><td>L-Glutamine (200 mM)</td><td>Gibco</td><td>25030081</td><td></td></tr><tr><td>Liberase T-Flex, Research Grade</td><td>Roche</td><td>5989132001</td><td></td></tr><tr><td>LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cells</td><td>Invitrogen</td><td>&nbsp;L3224</td><td></td></tr><tr><td>MATLAB software package</td><td>MathWorks</td><td><a href="https://www.mathworks.com/pricing-licensing.html?prodcode=ML">https://www.mathworks.com/pricing-licensing.html?prodcode=ML</a></td><td></td></tr><tr><td>NE-4000 Two Channel Programmable Syringe Pump</td><td>SyringePump</td><td>NE-4000-US</td><td></td></tr><tr><td>NEEDLE, HYPO STR DISP 27GX1/2"(100/BX) EXLINT</td><td>McKesson</td><td>408877</td><td></td></tr><tr><td>Nunc Cell Culture/Petri Dishes</td><td>Thermo-Scientific</td><td>174926</td><td></td></tr><tr><td>Olympus CKX41 Inverted Phase Contrast Microscope</td><td>Olympus</td><td>CKX41-B</td><td></td></tr><tr><td>PBS, pH 7.4</td><td>Gibco</td><td>10010023</td><td></td></tr><tr><td>Penicillin&sbquo; Streptomycin (10,000 U/mL)</td><td>Gibco</td><td>15140122</td><td></td></tr><tr><td>RPMI 1640</td><td>Gibco</td><td>11875093</td><td></td></tr><tr><td>RPMI 1640, no glucose</td><td>Gibco</td><td>11879020</td><td></td></tr><tr><td>Sodium Pyruvate (100 mM)</td><td>Gibco</td><td>11360070</td><td></td></tr><tr><td>Sodium Pyruvate (100 mM stock)</td><td>Gibco</td><td>11360070</td><td></td></tr><tr><td>Sorval ST 16R</td><td>Thermo-Scientific</td><td>75004381</td><td></td></tr><tr><td>Stainless steel 304 syringe needle</td><td>Milipore Sigma</td><td>Z117013-1EA</td><td></td></tr><tr><td>SURGICAL DRAPE, DISPOSABLE, 36 IN X 100 YD</td><td>Covetrus</td><td>10408</td><td></td></tr><tr><td>Surgical Scissors - ToughCut</td><td>Fine Science Tools</td><td>14054-13</td><td></td></tr><tr><td>Thumb Dressing Forceps 4.5" Serrated Delicate 1.3 mm Tip Width</td><td>Roboz</td><td>RS-8120</td><td></td></tr><tr><td>Thermo Scientific Nunc 200mL Centrifuge Tube</td><td>Thermo-Scientific</td><td>14-962-17</td><td></td></tr><tr><td>Thermo Scientific Nunc Centrifuge Bottle Adapters and Cushions</td><td>Thermo-Scientific</td><td>12-565-283B</td><td></td></tr><tr><td>TSDUB15 15L Dubnoff Shaking Water Bath</td><td>Fisher Scientific</td><td>300610298</td><td></td></tr><tr><td>TX-400 4 x 400 mL Swinging Bucket Rotor</td><td>Thermo-Scientific</td><td>75003181</td><td></td></tr><tr><td>Tygon S-50-HL</td><td>Tygon</td><td>AAXoooo3</td><td></td></tr><tr><td>Ultra Sensitive Rat Insulin ELISA kit</td><td>Crystal Chem</td><td>90060</td><td></td></tr></tbody></table>

isolatie en zuivering van het pancreaseiland door lewisratten met behulp van enzymatische vertering en dichtheidsgradiëntscheiding

Pancreatic Islet Isolation and Purification from Lewis Rats Using Enzymatic Digestion and Density-Gradient Separation

Isolatie en zuivering van het pancreaseiland door Lewisratten met behulp van enzymatische vertering en dichtheidsgradiëntscheiding

Pancreatic islets are essential for both clinical islet transplantation and preclinical studies aimed at understanding the mechanisms and progression of ...

pancreatic-islet-isolation-purification-from-lewis-rats-using

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Biology

Biologie

Het bereiken van hoge kwaliteit en de juiste hoeveelheid van de menselijke eilandjes is een van de prominente voorwaarden voor een succesvolle transplantatie eilandjes van Langerhans. In deze video, beschrijven we stap voor stap de procedures voor de menselijke eilandjes van Langerhans isolatie (deel I: de spijsvertering en het verzamelen van de pancreas weefsel) met behulp van een aangepaste automatische methode.

Human eilandjes van Langerhans Isolatie: Deel I: Spijsvertering en verzameling van alvleesklierweefsel

Het bereiken van hoge kwaliteit en de juiste hoeveelheid van de menselijke eilandjes is een van de prominente voorwaarden voor een succesvolle transplantatie eilandjes van Langerhans. In deze video, beschrijven we stap voor stap de procedures voor de menselijke eilandjes van Langerhans isolatie (deel II: zuivering en cultuur van de menselijke eilandjes) met behulp van een aangepaste automatische methode.

Human eilandjes van Langerhans Isolatie: Deel II: Zuivering en cultuur van de mens Eilandjes

Transplantatie van geïsoleerde eilandjes is voorgesteld om een ​​potentiële behandeling voor diabetes type 1 zijn. Hier beschrijven we een methode om de eilandjes van de muis pancreata isoleren en transplantatie ze aan de subcapsulaire ruimte van de nier.

Een methode voor Murine Islet Isolatie en subcapsulaire Niertransplantatie

Het aanbod van type 2 diabetes eilandjes voor onderzoek ontoereikend is. Hier delen we ons protocol voor het isoleren van eilandjes van patiënten die een gedeeltelijke pancreatectomy. Deze aanpak is een unieke locatie voor het verkrijgen van eilandjes van type 2 diabetes en klinisch afgestemd zonder diabetes in voldoende aantallen voor basis-en klinische studies.

Isolatie van de Mens Eilandjes van Gedeeltelijk Pancreatectomized Patiënten

Pancreatic islets, also called the Islets of Langerhans, are a cluster of endocrine cells which produces hormones for glucose regulation and other important biological functions. This protocol explains a method for isolating pancreatic islets by first perfusing collagenase via Ampulla of Vater to digest the exocrine pancreatic tissue and a simplified density gradient centrifugation to purify the islets from mice.

Pancreatic Islets Isolation through Collagenase Perfusion: An Enzymatic Method to Isolate Pancreatic Islet Cells

Cruciale uitdagingen voor het onderzoeksgebied diabetes zijn de moleculaire mechanismen die eilandje β-cellen replicatie en op werkwijzen voor het stimuleren van β-celregeneratie ontwikkelen begrijpen. Hierin een high-gehalte screening methode voor het identificeren en evalueren van de β-cel-replicatie bevorderende activiteit van kleine moleculen wordt gepresenteerd.

Een High-gehalte<em&gt; In Vitro</em&gt; Eilandjes van Langerhans β-cel Replication Discovery Platform

We beschrijven een protocol hierin voor het isoleren van intacte eilandjes van neonatale muizen. Pancreata werden gedeeltelijk gedigereerd met collagenase, gevolgd door wassen en handmatige oogst. 20-80 eilandje clusters verkrijgbaar per pancreas van pasgeboren muizen, die geschikt zijn voor verschillende eilandje studies.

Effectieve Isolatie van Functionele eilandjes van neonatale muis alvleesklier

Dit eilandje isolatie protocol beschreef een roman route van Collagenase injectie om het exocriene weefsel te verteren en een vereenvoudigde gradiënt procedure om de eilandjes van muizen te zuiveren. Het gaat om enzymatische spijsvertering, gradiënt scheiding/zuivering en eilandje hand-picking. Succesvolle isolatie kan 250 – 350 hoge kwaliteit en volledig functionele eilandjes per muis opleveren.

Een eenvoudig hoog rendement protocol voor isolatie van de alvleesklier eiland van muizen

Dit protocol biedt een stapsgewijze handleiding voor de aanschaf van een alvleesklier van varkens voor isolatie en zuivering van eilandjes.

Chirurgische perfusie en isolatie van de alvleesklier van varkens voor isolatie van eilandjes

Deze studie ontwikkelde een vereenvoudigde methode om efficiënt functionele primaire eilandjes en acinaire cellen uit de muizenalvleesklier te extraheren, wat een waardevol hulpmiddel biedt voor het bestuderen van intercellulaire communicatie in de pathogenese van Acute Pancreatitis-geïnduceerde Diabetes.

Een eenvoudige en snelle methode voor gelijktijdige isolatie van primaire eilandjes en primaire alvleesklier-acinaire cellen van muizen

Leer een gestandaardiseerd protocol voor de isolatie en zuivering van pancreaseilanden uit Lewis-ratten met behulp van enzymatische vertering en dichtheidsgradiëntscheiding. Deze methode optimaliseert opbrengst, zuiverheid en levensvatbaarheid voor diabetesonderzoek.

Isolatie en zuivering van het pancreaseiland door Lewisratten met behulp van enzymatische vertering en dichtheidsgradiëntscheiding (Video) | JoVE Journal

Isolatie en zuivering van het pancreaseiland door Lewisratten met behulp van enzymatische vertering en dichtheidsgradiëntscheiding

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