Name: a syngeneic pancreatic cancer mouse model to study the effects of irreversible electroporation
Uploaded: 2018-06-08T12:30:00
Description: Watch this Scientific Journal Video about A Syngeneic Pancreatic Cancer Mouse Model to Study the Effects of Irreversible Electroporation at JoVE.com

RRW har fått stöd för detta arbete från Collaborative translationell forskningsbidrag finansieras av San Diego C3 Padres pedalen den orsaken (#PTC2017).

Alla djurförsök utförs följande detta protokoll skall godkännas av respektive institutionella djur vård och användning kommittén (IACUC). Alla förfaranden beskrivs här har godkänts av IACUC UCSD.
1. upphandla mottagarens djur
Obs: KPC-Luc 4580 cell linjen inrättades från en tumör som uppstår i en LSL-KrasG12D / +; LSL-Trp53R172H / +; PDX1Cre / +; LSL-ROSA26 Luc / + mus (KPC), som är en genetiskt modifierad mo...

<ol>
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E., 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=Dynamics+of+the+immune+reaction+to+pancreatic+cancer+from+inception+to+invasion.">Dynamics of the immune reaction to pancreatic cancer from inception to invasion.</a> Cancer Res. 67 (19), 9518-9527 (2007).</li><li>Lee, E. W., Loh, C. T., Kee, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+guided+percutaneous+irreversible+electroporation:+ultrasound+and+immunohistological+correlation.">Imaging guided percutaneous irreversible electroporation: ultrasound and immunohistological correlation.</a> Technol Cancer Res Treat. 6 (4), 287-294 (2007).</li><li>Charpentier, K. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Irreversible+electroporation+for+the+ablation+of+liver+tumors:+are+we+there+yet?.">Irreversible electroporation for the ablation of liver tumors: are we there yet?.</a> Arch Surg. 147 (11), 1053-1061 (2012).</li><li>Narayanan, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Irreversible+Electroporation.">Irreversible Electroporation.</a> Seminars in Interventional Radiology. 32 (4), 349-355 (2015).</li><li>Martin, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+200+locally+advanced+(stage+III)+pancreatic+adenocarcinoma+patients+with+irreversible+electroporation:+safety+and+efficacy.">Treatment of 200 locally advanced (stage III) pancreatic adenocarcinoma patients with irreversible electroporation: safety and efficacy.</a> Ann Surg. 262 (3), 486-494 (2015).</li><li>Belfiore, M. 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=Percutaneous+CT-guided+irreversible+electroporation+followed+by+chemotherapy+as+a+novel+neoadjuvant+protocol+in+locally+advanced+pancreatic+cancer:+Our+preliminary+experience.">Percutaneous CT-guided irreversible electroporation followed by chemotherapy as a novel neoadjuvant protocol in locally advanced pancreatic cancer: Our preliminary experience.</a> Int J Surg. 21 Suppl 1, S34-S39 (2015).</li><li>Belfiore, G., 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=Concurrent+chemotherapy+alone+versus+irreversible+electroporation+followed+by+chemotherapy+on+survival+in+patients+with+locally+advanced+pancreatic+cancer.">Concurrent chemotherapy alone versus irreversible electroporation followed by chemotherapy on survival in patients with locally advanced pancreatic cancer.</a> Med Oncol. 34 (3), 38 (2017).</li><li>Chu, K. F., Dupuy, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermal+ablation+of+tumours:+biological+mechanisms+and+advances+in+therapy.">Thermal ablation of tumours: biological mechanisms and advances in therapy.</a> Nat Rev Cancer. 14 (3), 199-208 (2014).</li><li>Wissniowski, T. 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Y., 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=Activation+of+dendritic+cells+by+local+ablation+of+hepatocellular+carcinoma.">Activation of dendritic cells by local ablation of hepatocellular carcinoma.</a> J Hepatol. 43 (5), 817-822 (2005).</li><li>Fietta, A. M., 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=Systemic+inflammatory+response+and+downmodulation+of+peripheral+CD25+Foxp3++T-regulatory+cells+in+patients+undergoing+radiofrequency+thermal+ablation+for+lung+cancer.">Systemic inflammatory response and downmodulation of peripheral CD25+Foxp3+ T-regulatory cells in patients undergoing radiofrequency thermal ablation for lung cancer.</a> Hum Immunol. 70 (7), 477-486 (2009).</li><li>Jiang, C., Davalos, R. V., Bischof, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+basic+to+clinical+studies+of+irreversible+electroporation+therapy.">A review of basic to clinical studies of irreversible electroporation therapy.</a> IEEE Trans Biomed Eng. 62 (1), 4-20 (2015).</li><li>Neal, R. E., 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=Improved+local+and+systemic+anti-tumor+efficacy+for+irreversible+electroporation+in+immunocompetent+versus+immunodeficient+mice.">Improved local and systemic anti-tumor efficacy for irreversible electroporation in immunocompetent versus immunodeficient mice.</a> PLoS One. 8 (5), e64559 (2013).</li><li>Hingorani, S. R., 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=Trp53R172H+and+KrasG12D+cooperate+to+promote+chromosomal+instability+and+widely+metastatic+pancreatic+ductal+adenocarcinoma+in+mice.">Trp53R172H and KrasG12D cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice.</a> Cancer Cell. 7 (5), 469-483 (2005).</li><li>Safran, M., 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=Mouse+reporter+strain+for+noninvasive+bioluminescent+imaging+of+cells+that+have+undergone+Cre-mediated+recombination.">Mouse reporter strain for noninvasive bioluminescent imaging of cells that have undergone Cre-mediated recombination.</a> Mol Imaging. 2 (4), 297-302 (2003).</li><li>Martin, R. C., McFarland, K., Ellis, S., Velanovich, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Irreversible+electroporation+in+locally+advanced+pancreatic+cancer:+potential+improved+overall+survival.">Irreversible electroporation in locally advanced pancreatic cancer: potential improved overall survival.</a> Ann Surg Oncol. 20 Suppl 3, S443-S449 (2013).</li><li>Neal, R. E., Garcia, P. A., Robertson, J. L., Davalos, R. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+Characterization+and+Numerical+Modeling+of+Tissue+Electrical+Conductivity+during+Pulsed+Electric+Fields+for+Irreversible+Electroporation+Treatment+Planning.">Experimental Characterization and Numerical Modeling of Tissue Electrical Conductivity during Pulsed Electric Fields for Irreversible Electroporation Treatment Planning.</a> IEEE Transactions on Biomedical Engineering. 59 (4), 1076-1085 (2012).</li></ol>

I denna studie har vi visat en immunkompetenta musmodell för PC som kan användas för att studera effekterna av IRE tumör tillväxt. För närvarande, används IRE som en icke-termisk ablationsteknik endast i utvalda lokalt avancerad PC patienter som inte har avlägsna sjukdomsprogression efter månader av preoperativ terapi. Dess användning har därför begränsats eftersom de flesta patienter med lokalt avancerad PC utveckla avlägsen metastaserande sjukdom20. Denna modell kommer att tjäna ...

Pankreas duktal adenokarcinom (PC) väntas bli den näst vanligaste orsaken till dödsfall i cancer i USA runt 20201. Majoriteten av patienter som diagnostiserats med PC kommer att så småningom dö från avlägsna metastaserande sjukdom2. Den PC närmiljön är notoriskt immunsuppressiva och chemoresistant. Dess Desmoplastiskt stroma innehåller en brist på effektor (anti-tumör) T-celler och en framträdande av immunsuppressiva leukocyter, inklusive tumör-associerad makrofager (TAMs), myeloisk härledda suppressor celler (MDSCs) och regulatoriska T-celler (Tregs)3 . Dessa ligger bakom behovet av att utveckla multimodala strategier för att motverkar dessa effekter av mikromiljö.
IRE har utvecklats som en icke-termisk metod av tumör ablation. Till skillnad från termiska ablation tekniker, IRE orsaka inte snabba coagulative nekros men istället resulterar i gradvis apoptotiska cell död4. Huvudsakligen för Pankreastumör, IRE är inte sårbara ”kylfläns” effekter och kan utföras intill blodkärlen5. Denna teknik har 510(k) clearance från FDA6 och för närvarande används kliniskt, för utvalda patienter med lokalt avancerad eller borderline resektabel pankreascancer. I den största publicera serien av IRE för PC7var medianvärdet för överlevnad för patienter som genomgår IRE ungefär dubbel överlevnaden hos patienter som behandlats med moderna kemoterapi ensam utan resektion8,9.
Flera studier har visat att termisk ablation inducerar en systemisk immunsvar i andra tumörtyper (recenserade i Chu o.a. 10). radiofrekvent ablation (RFA) i djurens tumör modeller leder till ökad T cell infiltrerar11,12, inklusive en ökning i aktiverade naturliga mördarceller (NK) i Hepatocellulär cancer patienter13, 14, och en minskning av immunsuppressiva Tregs i lung cancer patienter15. Ett mycket mindre antal studier har undersökt immun, microenvironmental, och skada Svaren till IRE16. IRE har visat sig stimulera en systemisk immunsvar i immunkompetenta musmodeller där tillväxten av sekundära (kontralateral) renal cell organtransplantationer var minskas eller förhindras genom IRE av en primär tumör två veckor tidigare17. De påpekade också att immunkompetenta möss krävs mindre spänning för fullständig regression än gjorde immunsupprimerade möss. Det har varit hypotesen att IRE kan resultera i förbättrad antigen-presentation jämfört med den coagulative nekrosen av termiska ablation, men detta har inte specifikt studerats.
Vi har utvecklat en syngena musmodell av PC från KPC-Luc 4580 cell linje (gåva från J.J. Yeh vid University of North Carolina), som härrör från en tumör som utvecklats i en manlig LSL-KrasG12D / +; LSL-Trp53R172H / +; PDX1Cre / +; LSL-ROSA26 Luc / + mus, att studera lokala och systemiska effekter av IRE18,19. Detta luciferas-uttryckande cellinje är immunogena och också tumörframkallande hos immunkompetenta C57BL/6 möss när injiceras SQ eller orthotopically och pålitligt producerar levermetastaser när injiceras i mjälten. Vi har använt en programmerbar fyrkantsvåg pulsgenerator för att leverera 100-µs pulser av el i en spänning eller distans förhållande av 1 500 V/cm med en två-nål array sonden (avgränsade med 5 mm) eller platinum tweezer-derna till SQ eller ortotop tumörer, respektive i möss att modellera effekterna av IRE i ett litet djur.

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			<td height="40" style="height:40px;width:216px;">ECM 830 square wave electroporator</td>
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			<td height="40" style="height:40px;">2 needle array electrode</td>
			<td>Harvard Apparatus</td>
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			<td></td>
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			<td height="40" style="height:40px;">Safety foot switch</td>
			<td>Harvard Apparatus</td>
			<td>45-0211</td>
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			<td height="40" style="height:40px;">Platinum Tweezer-trode</td>
			<td>Harvard Apparatus</td>
			<td>&#160;45-0486</td>
			<td></td>
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			<td height="40" style="height:40px;">DMEM-F12 media</td>
			<td>ThermoFisher Scientific</td>
			<td>11320-033</td>
			<td></td>
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			<td height="40" style="height:40px;">Fetal Bovine Serum</td>
			<td>ThermoFisher Scientific</td>
			<td>10437028</td>
			<td></td>
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			<td height="40" style="height:40px;">Trypsin</td>
			<td>ThermoFisher Scientific</td>
			<td>25200056</td>
			<td></td>
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			<td height="40" style="height:40px;">Matrigel</td>
			<td>Corning&#160;</td>
			<td>354230</td>
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			<td height="40" style="height:40px;">Isoflurane</td>
			<td>Sigma-Aldrich, Inc.</td>
			<td>792632</td>
			<td></td>
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			<td height="40" style="height:40px;">Lacrilube</td>
			<td>Fisher Scientific&#160;</td>
			<td>19090646</td>
			<td></td>
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			<td height="40" style="height:40px;">Buprenorphine</td>
			<td>Fisher Scientific&#160;</td>
			<td>NC1292810</td>
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			<td height="40" style="height:40px;">D-luciferrin</td>
			<td>Perkin Elmer</td>
			<td>122799</td>
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			<td height="40" style="height:40px;">IVIS&#160; Spectrum In Vivo Imaging System</td>
			<td>Perkin Elmer</td>
			<td>124262</td>
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			<td height="40" style="height:40px;">Mouse strain C57BL/6J</td>
			<td>The Jackson Laboratory&#160;</td>
			<td>000664/Black 6</td>
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			<td height="40" style="height:40px;">Cell line (KPC-Luc 4580)</td>
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			<td height="40" style="height:40px;">BD Precisionglide syringe needles</td>
			<td>Sigma-Aldrich, Inc.</td>
			<td>Z192406</td>
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			<td height="40" style="height:40px;">Alcohol Swab(70% isopropyl alcohol )</td>
			<td>BD</td>
			<td>326895</td>
			<td></td>
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			<td height="40" style="height:40px;">Digital calipers</td>
			<td>ThermoFisher Scientific</td>
			<td>14-648-17</td>
			<td></td>
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			<td height="40" style="height:40px;">Disposable Scalpels, Sterile</td>
			<td>VWR</td>
			<td>21909</td>
			<td></td>
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			<td height="40" style="height:40px;">Cotton Tipped Applicators</td>
			<td>VWR</td>
			<td>89198</td>
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			<td height="40" style="height:40px;">Suture Needle, 45 cm, Size 6-0</td>
			<td>Harvard Apparatus</td>
			<td>72-3308</td>
			<td></td>
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			<td height="21" style="height:21px;">Suture Needle, 45 cm, Size 4-0</td>
			<td>Harvard Apparatus</td>
			<td>72-3314</td>
			<td></td>
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			<td height="21" style="height:21px;">Povidone-iodine 10%</td>
			<td>BD</td>
			<td>29900-404</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Disposable Warming Pad&#160;</td>
			<td>KENT SCIENTIFIC CORP.</td>
			<td>TP-3E</td>
			<td></td>
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			<td height="21" style="height:21px;">Mouse Hair Clipper</td>
			<td>KENT SCIENTIFIC CORP.</td>
			<td>CL8787</td>
			<td></td>
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			<td height="21" style="height:21px;">Surgical Drape</td>
			<td>Harvard Apparatus</td>
			<td>59-7421</td>
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			<td height="21" style="height:21px;">Phosphate-buffered Saline</td>
			<td>ThermoFisher Scientific</td>
			<td>10010023</td>
			<td></td>
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a syngeneic pancreatic cancer mouse model to study the effects of irreversible electroporation

Pankreascancer (PC), en sjukdom som dödar cirka 40 000 patienter varje år i USA, har framgångsrikt undvikit flera behandlingsmetoder inklusive de lovande immunoterapeutisk strategierna. Irreversibel elektroporation (IRE) är en icke-termisk ablationsteknik som inducerar tumörcelldöd utan förstörelse av intilliggande kollagena strukturer, vilket gör att förfarandet som skall utföras i tumörer mycket nära blodkärl. Till skillnad från termiska ablation tekniker, IRE resulterar i gradvis apoptotisk celldöd, tillsammans med omedelbara ablation inducerad nekros, och är för närvarande i klinisk användning för utvalda patienter med lokalt avancerad PC. En ablativ, icke-målorganismer särskilt förfarande som IRE kan inducera en myriad av svaren i den tumör mikromiljö. Ett fåtal studier har tagit upp effekterna av IRE på tumörtillväxt i andra tumörtyper, men ingen har fokuserat på PC. Vi har utvecklat en syngena musmodell av PC där subkutan (SQ) och ortotop tumörer kan behandlas framgångsrikt med IRE i en mycket kontrollerad miljö, underlätta olika longitudinella studier efter proceduren. Denna djurmodell fungerar som ett robust system för att studera effekterna av IRE och sätt att förbättra den kliniska effekten av IRE.

Vi följde förfaranden som beskrivs ovan och genererade SQ tumörer på 5-6 vecka gamla vildtyp C57BL/6 möss inokuleras med 1 x 106 celler med 50% BMM. När storleken på tumören nått 5-6 mm i diameter, några av mössen var euthanized, deras tumörer var censurerade och implanteras orthotopically hos en mottagande C57BL/6 möss. IRE var utfört 10 dagar post implantation som visas i figur 1tidslinje. IRE utfördes på återstående möss med S...

Watch this Scientific Journal Video about A Syngeneic Pancreatic Cancer Mouse Model to Study the Effects of Irreversible Electroporation at JoVE.com

A Syngeneic Pancreatic Cancer Mouse Model to Study the Effects of Irreversible Electroporation

En syngena pankreascancer musmodell att studera effekterna av irreversibel elektroporation

Irreversibel elektroporation (IRE) är en icke-termisk ablationsteknik används för behandling av lokalt avancerad pankreascancer. Att vara en relativt ny teknik, är effekterna av IRE på tumörtillväxt dåligt känd. Vi har utvecklat en syngena musmodell som underlättar studera effekterna av IRE på pankreascancer.

Pancreatic cancer (PC), a disease which kills approximately 40,000 patients each year in the US, has successfully evaded several therapeutic approaches ...

The overall goal of this model is to study the effects of irreversible electroporation or IRE, on pancreatic cancer in immunocompetent mice. This model can help answer many key questions regarding the role of IRE in pancreatic cancer. Such as, how does IRE affect the local tumor microenvironment?The main advantage of this model is that it facilitate accurate tumor growth assessment in an immunocompetent system, without requiring knockout or mutant mouse models. This method can provide insight into the effects of IRE on the immune system, in an immunocompetent mouse model of pancreatic cancer. For subcutaneous tumor induction, first, shave the injection site on an anesthetized six-week old, male, C57 black six mouse, and use an alcohol swab to clean the site two times.Next, load a cold one-milliliter syringe with one to two times ten to the seven cells per milliliter and 250 microliters of basement membrane matrix. Attach a 25-gauge needle to the syringe, and load the needle with the tumor cell suspension. Press and retract the plunger a few times to remove any air bubbles.Carefully insert the needle under the skin of the injection site, at a 30 degree angle. With the needle parallel to the body, slowly inject 50 microliters of the tumor cells. After 10 seconds, slowly withdraw the needle, keeping it parallel without any sideways movement, and gently clean the injection site with an alcohol swab, before returning the mouse to its cage.When the tumor reaches five to seven millimeters, use sterile forceps and micro scissors to excise the subcutaneous tumor, under sterile conditions in a biosafety level two hood, taking care not to puncture the peritoneal cavity. Immediately wash the excised tumor, two times, in sterile PBS. And use sterile scalpels to mince the tumor into one to two cubed millimeter pieces.Then, transfer the tumor pieces into a Petri dish, containing cold, sterile DMEM basal medium on ice. To implant the tumor fragments, after confirming a lack of response to toe pinch, apply ointment to the anesthetized recipient animal's eyes, and place the mouse on its right side, so that the left side of the abdomen is exposed. Remove the abdominal hair with depilatory cream, and remove the cream with gauze.After disinfecting the exposed skin with three sequential 10%povidone plus iodine and alcohol wipes, use a sterile scalpel to make a 1.5 centimeter incision in the skin, one centimeter to the left of the midline, below the rib cage, and slightly medial to the spleen. Gently externalize the spleen with flat-tipped forceps and a sterile cotton applicator, and use the forceps to laterally retract the tail of the pancreas, at the bottom of the spleen. Using sterile forceps, grasp one finely-minced tumor piece, and pass a fine 6-O sterile suturing needle through the piece.Insert the needle containing tumor fragment into the tail of the pancreas, passing the suture slowly through the tissue. Take care to avoid contact between the tumor and the abdominal wall musculature and peritoneum, to prevent tumor seeding. When the tumor fragment has been secured, completely remove the needle from the tissue, and keep the organs completely externalized, for an additional 60 seconds, while inspecting for any signs of hemorrhage.If no bleeding is observed, use blunt forceps, or a cotton tip applicator, to gently return the spleen, pancreas, and tumor to the abdominal cavity. Before the tumor reaches six millimeters in diameter, place the anesthetized animal on its right side to allow access to the tumor, and use clippers and an alcohol swab to clean the tumor site. Using forceps, elevate the skin directly under the tumor, and insert two needle array electrodes through the skin, parallel to the body, to bracket the tumor, taking care that the electrodes do not penetrate the peritoneal cavity.Set the electroporator to deliver 100 microsecond pulses at a one hertz frequency, at 1500 volts per centimeter, for a total of 150 pulses, and use the foot pedal to deliver the pulses, separating each set of 10 pulses by 10 seconds, to allow heat dissipation between each set of pulses. As muscle twitching occurs during the IRE delivery, and is more pronounced during the initial set of pulses, ensure that the electrodes are still in the desired location, and that the animal is still sedated after each set of pulses. After no more than 200 seconds of dosing, remove the electrodes and record the actual voltage delivered, as displayed on the electroporator.Then, return the mouse to its cage. For IRE of an orthotopic tumor, eight to 10 days post tumor implantation, use blunt-nosed forceps to externalize the tumor with the forceps, and tightly capture the tumor with the platinum electrodes of the Tweezertrode, deliver the electroporation pulses, as just demonstrated, keeping the tumor externalized for the last set of pulses, for at least 60 seconds, to monitor for any signs of hemorrhage. Then, return the tumor to the abdominal cavity and close the incision, as demonstrated, monitoring the animal until it has fully recovered.In this representative experiment, subcutaneous tumors, in nearly 50%of the mice, regressed completely after 150 IRE pulses, with only minor regression after 75 pulses. Histological analysis of the tumor tissue, one week post IRE, revealed large regions of central tumor necrosis, that were not observed in control, untreated tumors, and that were flanked by live tumor tissue, in cases of incomplete tumor regression. Monitoring implanted orthotopic tumors growth rates revealed a consistent tumor growth over time in recipient animals, that could also be disrupted by IRE, demonstrating the ability of this model to simulate the effects of IRE, in an immunocompetent mouse model.Once mastered, this technique can be completed in 15 minutes per mouse, if it is performed properly. While attempting this procedure, it is important to remember to keep the tumor size within the working distance of the IRE electrodes available, as larger tumors will produce inconsistent responses due to incomplete ablation. Other treatment approaches, such as radiation or chemotherapies, can also be used to answer additional questions about whether combining treatments can improve tumor responses to IRE.After watching this video, you should have a good understanding of how to establish subcutaneous and orthotopic pancreatic cancers in mice, and how to perform IRE on these tumors.

Research

JoVE Journal

Cancer Research

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