Name: pancreatic duct infusion: an effective and selective method of drug and viral delivery
Uploaded: 2021-09-30T13:00:00
Description: Watch this Scientific Journal Video about Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery at JoVE.com

The authors have no acknowledgements. Funding was received from the Foundation of Health and Family Planning Commission of Zhejiang Province (Grant 20146242) and the Foundation of Wenzhou Science &amp; Technology Bureau (Grant Y20140248).

All animal experiments were approved by the Animal Research and Care Committee at the Children&#39;s Hospital of Pittsburgh and the University of Pittsburgh Institutional Animal Care and Use Committee.
1. Preoperative Preparation
<ol>
	<li>Administer inhaled isoflurane (1 - 3% for maintenance and up to 5% for induction) via a nose cone to an appropriate anesthetic level. Monitor the adequacy of anesthesia by toe pinch with forceps. The lack of a response is considered adequate, and be sure n...

<ol>
	<li>Gittes, G. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+biology+of+the+pancreas:+a+comprehensive+review.">Developmental biology of the pancreas: a comprehensive review.</a> Dev Biol. 326 (1), 4-35 (2009).</li><li>Gu, G., Dubauskaite, J., Melton, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+evidence+for+the+pancreatic+lineage:+NGN3++cells+are+islet+progenitors+and+are+distinct+from+duct+progenitors.">Direct evidence for the pancreatic lineage: NGN3+ cells are islet progenitors and are distinct from duct progenitors.</a> Development. 129 (10), 2447-2457 (2002).</li><li>Weinberg, N., Ouziel-Yahalom, L., Knoller, S., Efrat, S., Dor, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lineage+tracing+evidence+for+in+vitro+dedifferentiation+but+rare+proliferation+of+mouse+pancreatic+beta-cells.">Lineage tracing evidence for in vitro dedifferentiation but rare proliferation of mouse pancreatic beta-cells.</a> Diabetes. 56 (5), 1299-1304 (2007).</li><li>Raper, S. E., DeMatteo, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adenovirus-mediated+in+vivo+gene+transfer+and+expression+in+normal+rat+pancreas.">Adenovirus-mediated in vivo gene transfer and expression in normal rat pancreas.</a> Pancreas. 12 (4), 401-410 (1996).</li><li>Xiao, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pancreatic+cell+tracing,+lineage+tagging+and+targeted+genetic+manipulations+in+multiple+cell+types+using+pancreatic+ductal+infusion+of+adeno-associated+viral+vectors+and/or+cell-tagging+dyes.">Pancreatic cell tracing, lineage tagging and targeted genetic manipulations in multiple cell types using pancreatic ductal infusion of adeno-associated viral vectors and/or cell-tagging dyes.</a> Nat Protoc. 9 (12), 2719-2724 (2014).</li><li>Song, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EGFR+signaling+regulates+beta+cell+proliferation+in+adult+mice.">EGFR signaling regulates beta cell proliferation in adult mice.</a> J Biol Chem. 291 (43), (2016).</li><li>Xiao, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=M2+macrophages+promote+beta-cell+proliferation+by+up-regulation+of+SMAD7.">M2 macrophages promote beta-cell proliferation by up-regulation of SMAD7.</a> Proc Natl Acad Sci U S A. 111 (13), E1211-E1220 (2014).</li><li>Laukkarinen, J. M., Van Acker, G. J., Weiss, E. R., Steer, M. L., Perides, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+model+of+acute+biliary+pancreatitis+induced+by+retrograde+pancreatic+duct+infusion+of+Na-taurocholate.">A mouse model of acute biliary pancreatitis induced by retrograde pancreatic duct infusion of Na-taurocholate.</a> Gut. 56 (11), 1590-1598 (2007).</li><li>Lichtenstein, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+lung+injury+in+two+experimental+models+of+acute+pancreatitis:+infusion+of+saline+or+sodium+taurocholate+into+the+pancreatic+duct.">Acute lung injury in two experimental models of acute pancreatitis: infusion of saline or sodium taurocholate into the pancreatic duct.</a> Crit Care Med. 28 (5), 1497-1502 (2000).</li><li>Perides, G., van Acker, G. J., Laukkarinen, J. M., Steer, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+acute+biliary+pancreatitis+induced+by+retrograde+infusion+of+bile+acids+into+the+mouse+pancreatic+duct.">Experimental acute biliary pancreatitis induced by retrograde infusion of bile acids into the mouse pancreatic duct.</a> Nat Protoc. 5 (2), 335-341 (2010).</li><li>Guo, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specific+transduction+and+labeling+of+pancreatic+ducts+by+targeted+recombinant+viral+infusion+into+mouse+pancreatic+ducts.">Specific transduction and labeling of pancreatic ducts by targeted recombinant viral infusion into mouse pancreatic ducts.</a> Lab Invest. 93 (11), 1241-1253 (2013).</li><li>Xiao, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurogenin3+activation+is+not+sufficient+to+direct+duct-to-beta+cell+transdifferentiation+in+the+adult+pancreas.">Neurogenin3 activation is not sufficient to direct duct-to-beta cell transdifferentiation in the adult pancreas.</a> J Biol Chem. 288 (35), 25297-25308 (2013).</li></ol>

This work describes in detail the methodology behind pancreatic duct infusion, an effective mouse model for the delivery of genes and other molecules specifically and effectively to the pancreas. Ambiguity in the specifics of the procedure between various groups highlighted the need for a standardized protocol8,9,10.
There are several critical steps of the procedure, beginning with the selection of yo...

The pancreas is a bifunctional organ, with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, pancreatitis, and pancreatic cancer1. All three of these diseases mark active areas of study, not only to develop immediate therapy, but also to better understand their pathophysiology.
A targeted therapeutic delivery method would be beneficial. We have developed a pancreatic duct infusion technique that is an effective and selective method for drug and viral delivery to pancreatic cells. In this model, the pancreatic duct is selectively cannulated, and the common bile duct is occluded, allowing for delivery exclusively to the pancreas.
This technique can allow for the lineage tracing of specific cell types to further elucidate the developmental pathways important to pancreatic development and pathology2,3. Different promoters in viral vectors allow for the specific targeting of cell lines and for the introduction of genes into these cell lines4,5. This work demonstrates the expression of a green fluorescent protein (GFP) within the pancreas after the pancreatic duct infusion of a viral vector expressing GFP versus a sham surgery.

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			<td height="42" style="height:42px;width:288px;">Mice (25-30g, 8-12 weeks old)</td>
			<td style="width:171px;">Jackson Laboratory</td>
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			<td style="width:167px;"></td>
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			<td height="42" style="height:42px;width:288px;">Ketofen</td>
			<td style="width:171px;">Henry Schein Inc.</td>
			<td style="width:137px;">5487</td>
			<td></td>
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			<td height="42" style="height:42px;width:288px;">Ethanol (70%)</td>
			<td style="width:171px;">Sigma-Aldrich</td>
			<td style="width:137px;">34923</td>
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		<tr height="63">
			<td height="63" style="height:63px;width:288px;">Sterile Saline Solution</td>
			<td style="width:171px;">Baxter Healthcare Corp.</td>
			<td style="width:137px;">2B-13-00</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Protective Equipment</td>
			<td style="width:171px;"></td>
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			<td height="21" style="height:21px;width:288px;">Hair Removal Product</td>
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			<td>Nair or trimmer</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:288px;">Gauze Pads 2in x 2in</td>
			<td style="width:171px;">Fisher Healthcare</td>
			<td style="width:137px;">22-362-178</td>
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		<tr height="63">
			<td height="63" style="height:63px;width:288px;">Needles (30.5G and 25G)</td>
			<td style="width:171px;">Becton Dicksinson and Co.</td>
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		<tr height="63">
			<td height="63" style="height:63px;width:288px;">Syringe for Ketofen and Saline</td>
			<td style="width:171px;">Becton Dicksinson and Co.</td>
			<td style="width:137px;">309657</td>
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			<td height="42" style="height:42px;width:288px;">Syringe for Pump</td>
			<td style="width:171px;">Henke Sass Wolf</td>
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		<tr height="63">
			<td height="63" style="height:63px;width:288px;">Infusion Pump</td>
			<td style="width:171px;">Pump Systems Inc.</td>
			<td style="width:137px;">NE-1000</td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:288px;">Infusion Catheter</td>
			<td style="width:171px;">World Precision Instruments</td>
			<td style="width:137px;">CMF31G</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Curved Bulldog Clamps (2)</td>
			<td style="width:171px;">Roboz</td>
			<td style="width:137px;">RS-7439</td>
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			<td height="21" style="height:21px;width:288px;">Arruga Forceps</td>
			<td style="width:171px;">Roboz</td>
			<td style="width:137px;">RS-5163</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Adson Forcep</td>
			<td style="width:171px;">Roboz</td>
			<td style="width:137px;">RS-5234</td>
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		<tr height="24">
			<td height="24" style="height:24px;width:288px;">Needle Holder</td>
			<td style="width:171px;">Roboz</td>
			<td>RS-7882</td>
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			<td height="24" style="height:24px;width:288px;">Scissor</td>
			<td style="width:171px;">Roboz</td>
			<td>RS-5982</td>
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		<tr height="63">
			<td height="63" style="height:63px;width:288px;">Cotton Tip Applicators</td>
			<td style="width:171px;">Physician Sales and Services</td>
			<td>22-9988</td>
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			<td height="21" style="height:21px;width:288px;">Dissecting Microscope</td>
			<td style="width:171px;"></td>
			<td style="width:137px;"></td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:288px;">Suture</td>
			<td style="width:171px;">Owens and Minor</td>
			<td>SXMD1B402</td>
			<td></td>
		</tr>
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pancreatic duct infusion: an effective and selective method of drug and viral delivery

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, pancreatitis, and pancreatic cancer. All three of these diseases mark active areas of study, not only to develop immediate therapy, but also to better understand their pathophysiology. There are few tools to further these areas of study. Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. The technique requires the intricate dissection of the second portion of the duodenum and ampulla, followed by the occlusion of the bile duct and the cannulation of the pancreatic duct. Although the technique is technically challenging at first, the applications are myriad. Ambiguity in the specifics of the procedure between groups highlighted the need for a standard protocol. This work describes the expression of a green fluorescent protein (GFP) within the pancreas after the pancreatic duct infusion of a viral vector expressing GFP versus a sham surgery. The infusion and therefore expression is specific to the pancreas, without expression present in any other tissue type.

With practice and careful surgical technique, the survival rate of these mice should be greater than 95%. One week after infusion, the desired effect should be evident in the pancreas. Mice at 10 to 12 weeks of age were utilized for pancreatic duct infusions. Here we use an adeno-associated virus serotype 8 (AAV8) with a CMV promoter to express green fluorescent protein (GFP), as compared to a sham surgery. Mice pancreases were harvested 7 days after the infusions. Sections of pancreas, s...

Watch this Scientific Journal Video about Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery at JoVE.com

Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery

Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. A pancreatic duct infusion technique for drug and viral delivery to pancreatic cells is presented here.

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, ...

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