Name: a murine tail lymphedema model
Uploaded: 2021-02-10T13:00:00
Description: Watch this Scientific Journal Video about A Murine Tail Lymphedema Model at JoVE.com

Dette arbejde blev støttet af tilskud finansiering fra American Association of Plastic Surgeons Academic Scholarship og Department of Defense W81XWH2110135   til AHH. Æstetisk Kirurgi Uddannelse og Research Foundation tilskud til MS NIH U01DK119099, R01NS042617 og R01DK125835 til CKS.

Protokollen følger retningslinjerne fra institutionens etiske komité for dyreforsøg. Alle dyreforsøg blev godkendt af Indiana University School of Medicine Institutional Animal Care and Use Committee. Dyr blev opstaldet under en 12-timers lys-mørk cyklus med mad og vand ad libitum.
1. Kirurgisk forstyrrelse af musehale lymfeknuder
<ol><li>Brug otte uger gamle C57BL/6 mus med lige kønsfordeling.</li>
	<li>Placer en mus under generel anæstesi i et induktionskammer med 3-4% isoflurane i ...

<ol>
	<li>Kataru, R. 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=Fibrosis+and+secondary+lymphedema:+chicken+or+egg.">Fibrosis and secondary lymphedema: chicken or egg.</a> Translation Research. 209, 68-76 (2019).</li><li>Brayton, K. 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=Lymphedema+prevalence+and+treatment+benefits+in+cancer:+impact+of+a+therapeutic+intervention+on+health+outcomes+and+costs.">Lymphedema prevalence and treatment benefits in cancer: impact of a therapeutic intervention on health outcomes and costs.</a> PLoS One. 9 (12), 114597 (2014).</li><li>Mendoza, N., Li, A., Gill, A., Tyring, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Filariasis:+diagnosis+and+treatment.">Filariasis: diagnosis and treatment.</a> Dermatology and Therapy. 22 (6), 475-490 (2009).</li><li>Rockson, S. G., Rivera, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimating+the+population+burden+of+lymphedema.">Estimating the population burden of lymphedema.</a> Annals of the New York Academy of Sciences. 1131, 147-154 (2008).</li><li>Soran, 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=Breast+cancer-related+lymphedema--what+are+the+significant+predictors+and+how+they+affect+the+severity+of+lymphedema.">Breast cancer-related lymphedema--what are the significant predictors and how they affect the severity of lymphedema.</a> Breast Journal. 12 (6), 536-543 (2006).</li><li>Hayes, S. C., 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=Upper-body+morbidity+after+breast+cancer:+incidence+and+evidence+for+evaluation,+prevention,+and+management+within+a+prospective+surveillance+model+of+care.">Upper-body morbidity after breast cancer: incidence and evidence for evaluation, prevention, and management within a prospective surveillance model of care.</a> Cancer. 118, 2237-2249 (2012).</li><li>Carl, H. 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=Systematic+Review+of+the+Surgical+Treatment+of+Extremity+Lymphedema.">Systematic Review of the Surgical Treatment of Extremity Lymphedema.</a> Journal of Reconstructive Microsurgery. 33 (6), 412-425 (2017).</li><li>Garza, R., Skoracki, R., Hock, K., Povoski, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comprehensive+overview+on+the+surgical+management+of+secondary+lymphedema+of+the+upper+and+lower+extremities+related+to+prior+oncologic+therapies.">A comprehensive overview on the surgical management of secondary lymphedema of the upper and lower extremities related to prior oncologic therapies.</a> BMC Cancer. 17 (1), 468 (2017).</li><li>Hassanein, A. H., 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=Deep+Inferior+Epigastric+Artery+Vascularized+Lymph+Node+Transfer:+A+Simple+and+Safe+Option+for+Lymphedema.">Deep Inferior Epigastric Artery Vascularized Lymph Node Transfer: A Simple and Safe Option for Lymphedema.</a> Journal of Plastic, Reconstructive, Aesthetic Surgery. 73 (10), 1897-1916 (2020).</li><li>Hassanein, A. H., Sacks, J. M., Cooney, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimizing+perioperative+lymphatic-venous+anastomosis+localization+using+transcutaneous+vein+illumination,+isosulfan+blue,+and+indocyanine+green+lymphangiography.">Optimizing perioperative lymphatic-venous anastomosis localization using transcutaneous vein illumination, isosulfan blue, and indocyanine green lymphangiography.</a> Microsurgery. 37 (8), 956-957 (2017).</li><li>Chang, D. W., Masia, J., Garza, R., Skoracki, R., Neligan, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymphedema:+Surgical+and+Medical+Therapy.">Lymphedema: Surgical and Medical Therapy.</a> Plastic and Reconstructive Surgery. 138, 209-218 (2016).</li><li>Gould, D. J., Mehrara, B. J., Neligan, P., Cheng, M. H., Patel, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymph+node+transplantation+for+the+treatment+of+lymphedema.">Lymph node transplantation for the treatment of lymphedema.</a> Journal of Surgical Oncology. 118 (5), 736-742 (2018).</li><li>Cook, J. 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=Immediate+Lymphatic+Reconstruction+after+Axillary+Lymphadenectomy:+A+Single-Institution+Early+Experience.">Immediate Lymphatic Reconstruction after Axillary Lymphadenectomy: A Single-Institution Early Experience.</a> Annals of Surgical Oncology. , (2020).</li><li>Cook, J. A., Hassanein, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ASO+Author+Reflections:+Immediate+Lymphatic+Reconstruction:+A+Proactive+Approach+to+Breast+Cancer-Related+Lymphedema.">ASO Author Reflections: Immediate Lymphatic Reconstruction: A Proactive Approach to Breast Cancer-Related Lymphedema.</a> Annals of Surgical Oncology. , (2020).</li><li>Johansson, K., Branje, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arm+lymphoedema+in+a+cohort+of+breast+cancer+survivors+10+years+after+diagnosis.">Arm lymphoedema in a cohort of breast cancer survivors 10 years after diagnosis.</a> Acta Oncologica. 49 (2), 166-173 (2010).</li><li>Johnson, A. 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=Lymphedema+Incidence+After+Axillary+Lymph+Node+Dissection:+Quantifying+the+Impact+of+Radiation+and+the+Lymphatic+Microsurgical+Preventive+Healing+Approach.">Lymphedema Incidence After Axillary Lymph Node Dissection: Quantifying the Impact of Radiation and the Lymphatic Microsurgical Preventive Healing Approach.</a> Annals of Plastic Surgery. 82, 234-241 (2019).</li><li>Gartner, R., Mejdahl, M. K., Andersen, K. G., Ewertz, M., Kroman, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+in+self-reported+arm-lymphedema+in+Danish+women+treated+for+early-stage+breast+cancer+in+2005+and+2006--a+nationwide+follow-up+study.">Development in self-reported arm-lymphedema in Danish women treated for early-stage breast cancer in 2005 and 2006--a nationwide follow-up study.</a> Breast. 23 (4), 445-452 (2014).</li><li>Shin, W. S., Rockson, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+the+molecular+and+mechanistic+study+of+lymphatic+biology+and+disease.">Animal models for the molecular and mechanistic study of lymphatic biology and disease.</a> Annals of the New York Academy of Sciences. 1131, 50-74 (2008).</li><li>Tobbia, D., 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=Lymphedema+development+and+lymphatic+function+following+lymph+node+excision+in+sheep.">Lymphedema development and lymphatic function following lymph node excision in sheep.</a> Journal of Vascular Research. 46 (5), 426-434 (2009).</li><li>Olszewski, W., Machowski, Z., Sokolowski, J., Nielubowicz, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+lymphedema+in+dogs.">Experimental lymphedema in dogs.</a> Journal of Cardiovascular Surgery. 9 (2), 178-183 (1968).</li><li>Rutkowski, J. M., Moya, M., Johannes, J., Goldman, J., Swartz, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secondary+lymphedema+in+the+mouse+tail:+Lymphatic+hyperplasia,+VEGF-C+upregulation,+and+the+protective+role+of+MMP-9.">Secondary lymphedema in the mouse tail: Lymphatic hyperplasia, VEGF-C upregulation, and the protective role of MMP-9.</a> Microvascular Research. 72 (3), 161-171 (2006).</li><li>Tabibiazar, 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=Inflammatory+manifestations+of+experimental+lymphatic+insufficiency.">Inflammatory manifestations of experimental lymphatic insufficiency.</a> PLoS Medicine. 3 (7), 254 (2006).</li><li>Slavin, S. A., Van den Abbeele, A. D., Losken, A., Swartz, M. A., Jain, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Return+of+lymphatic+function+after+flap+transfer+for+acute+lymphedema.">Return of lymphatic function after flap transfer for acute lymphedema.</a> Annals of Surgery. 229 (3), 421-427 (1999).</li><li>Zampell, J. C., 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=Toll-like+receptor+deficiency+worsens+inflammation+and+lymphedema+after+lymphatic+injury.">Toll-like receptor deficiency worsens inflammation and lymphedema after lymphatic injury.</a> American Journal of Physiology-Cell Physiology. 302 (4), 709-719 (2012).</li><li>Gardenier, J. C., 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=Diphtheria+toxin-mediated+ablation+of+lymphatic+endothelial+cells+results+in+progressive+lymphedema.">Diphtheria toxin-mediated ablation of lymphatic endothelial cells results in progressive lymphedema.</a> JCI Insight. 1 (15), 84095 (2016).</li><li>Weiler, M. J., Cribb, M. T., Nepiyushchikh, Z., Nelson, T. S., Dixon, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+mouse+tail+lymphedema+model+for+observing+lymphatic+pump+failure+during+lymphedema+development.">A novel mouse tail lymphedema model for observing lymphatic pump failure during lymphedema development.</a> Scientific Reports. 9 (1), 10405 (2019).</li><li>Avraham, T., 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=Th2+differentiation+is+necessary+for+soft+tissue+fibrosis+and+lymphatic+dysfunction+resulting+from+lymphedema.">Th2 differentiation is necessary for soft tissue fibrosis and lymphatic dysfunction resulting from lymphedema.</a> FASEB J. 27 (3), 1114-1126 (2013).</li><li>Zampell, J. C., 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=CD4(+)+cells+regulate+fibrosis+and+lymphangiogenesis+in+response+to+lymphatic+fluid+stasis.">CD4(+) cells regulate fibrosis and lymphangiogenesis in response to lymphatic fluid stasis.</a> PLoS One. 7 (11), 49940 (2012).</li><li>Arruda, G., Ariga, S., de Lima, T. M., Souza, H. P., Andrade, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+modified+mouse-tail+lymphedema+model.">A modified mouse-tail lymphedema model.</a> Lymphology. 53 (1), 29-37 (2020).</li><li>Jun, H., 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=Modified+Mouse+Models+of+Chronic+Secondary+Lymphedema:+Tail+and+Hind+Limb+Models.">Modified Mouse Models of Chronic Secondary Lymphedema: Tail and Hind Limb Models.</a> Annals of Vascular Surgery. 43, 288-295 (2017).</li><li>Karkkainen, M. J., 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=A+model+for+gene+therapy+of+human+hereditary+lymphedema.">A model for gene therapy of human hereditary lymphedema.</a> Proceedings of the National Academy of Sciences of the United States of America. 98 (22), 12677-12682 (2001).</li><li>Yoon, Y. S., 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=VEGF-C+gene+therapy+augments+postnatal+lymphangiogenesis+and+ameliorates+secondary+lymphedema.">VEGF-C gene therapy augments postnatal lymphangiogenesis and ameliorates secondary lymphedema.</a> Journal of Clinical Investigation. 111 (5), 717-725 (2003).</li><li>Gallego-Perez, D., 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=Topical+tissue+nano-transfection+mediates+non-viral+stroma+reprogramming+and+rescue.">Topical tissue nano-transfection mediates non-viral stroma reprogramming and rescue.</a> Nature Nanotechnology. 12 (10), 974-979 (2017).</li><li>Moore, J. 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Lymphedema er kategoriseret som en primær (medfødt) eller sekundær (iatrogen lymfe) skade38,39. Sekundær Lymphedema omfatter 99% aftilfældene 39. Sekundær lymphedema er oftest forårsaget af infektion (filariasis) eller post-onkologisk behandling med lymfektomi eller stråling4,39. En translationel dyremodel er udfordrende for sekundær Lymphedema, da 70% af dyr behandlet med...

Lymphedema er ekstremitet hævelse forårsaget af lymfe dysfunktion. Det berørte lem forstørrer på grund af ophobning af væske, fedt og fibrose1. Lymphedema påvirker 250 millioner mennesker på verdensplan2,3,4. Det anslås, at 20-40% af patienter, der gennemgår behandling for solide maligniteter, såsom brystkræft, melanom, gynækologiske / urologiske tumorer, eller sarkomer, udvikle Lymphedema2,4,5. Sygelighed fra Lymphedema omfatter tilbagevendende infektioner, smerter, og deformitet6. Der er ingen kur mod denne progressive, livslange sygdom. Nuværende behandlinger er variaby effektiv7 og omfatter kompression, komplet dekongetiv behandling af fysioterapeuter, excisional procedurer, og mikrokirurgiske operationer, herunder vaskulære lymfeknude overførsel og lymfeknude bypass7,8,9,10,11,12,13,14. Den ideelle behandling for Lymphedema er endnu ikke opdaget.
Undersøgelse af mekanisme og behandling af Lymphedema har været begrænset. Der er en gennemsnitlig forsinket debut på et år efter lymfeskade15,16 og de fleste personer, der oplever iatrogen fornærmelse med stråling og kirurgi ikke udvikler Lymphedema4,6,17. Selvom store dyremodeller, herunder hunde, får og gris er blevet beskrevet18,19,20, har musehalemodellen været den mest anvendte på grund af lethed, omkostninger og reproducibilty. Musemodeller til undersøgelse af Lymphedema omfatter en halemodel, difteri-toksinmedieret lymfeablation og aksillære eller popliteal lymfeknude dissektion21,22,23,24,25,26. De fleste hale modeller bruger en fokal, fuld tykkelse hud excision med lymfekanal klipning, der udføres nær bunden af halen22, hvilket resulterer i hale hævelse og histologiske træk svarende til human Lymphedema24,27,28,29. Standard murine halemodellen forsvinder dog typisk spontant på så få som 20 dage og ledsages af periodisk halenekrose30. Lymphedema mus hale model udvider en vedvarende Lymphedema ud over 15 uger, viser bekræftet arteriel og venøs patency, og giver funktionelle lymfe dysfunktion vurdering.
En murine hale model af Lymphedema giver mulighed for evaluering af nye terapeutiske til behandling af Lymphedema. Genbaserede strategier er blevet brugt i musemodellen medieret af virale vektorer31,32. Vi bruger også en ny væv nanotransfection teknologi (TNT) for genetisk last levering til lymfødem musehale. TNT letter direkte, transkutan genlevering ved hjælp af en chip med nanokanaler i et hurtigt fokuseret elektrisk felt33,34,35,36. Modellen omfatter brug af TNT2.0 at give mulighed for fokal gen levering af potentielle gen-baserede therapeutics til lymfeskade stedet for musen hale35.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Surgical Microscope</td>
			<td>Leica, Wetzlar, Germany</td>
			<td>MSV266</td>
			<td></td>
		</tr>
		<tr>
			<td>Adherent Dressing (Tegaderm)</td>
			<td>3M, St. Paul, Minn.</td>
			<td>1626W</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser speckle (Pericam PSI System )</td>
			<td>Perimed AB, Stockholm, Sweden)</td>
			<td>PSIZ</td>
			<td></td>
		</tr>
		<tr>
			<td>Near-infrared laser (LUNA)</td>
			<td>Stryker (Formerly Novadaq Technologies, Toronto, Canada)</td>
			<td>LU3000</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6 mice</td>
			<td>Jackson Laboratories</td>
			<td>000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-Adson Forceps - 1x2 Teeth</td>
			<td>Fine Science Tools (USA) Inc.</td>
			<td>11019-12</td>
			<td></td>
		</tr>
		<tr>
			<td>V-Hook</td>
			<td>Fine Science Tools (USA) Inc.</td>
			<td>18052-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel SS NO15</td>
			<td>Fischer Scientific</td>
			<td>29556</td>
			<td></td>
		</tr>
		<tr>
			<td>Disposable Needle 30GX1</td>
			<td>Fischer Scientific</td>
			<td>305128</td>
			<td></td>
		</tr>
		<tr>
			<td>Operating Scissors</td>
			<td>Fischer Scientific</td>
			<td>12-460-796</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgi-Or Jeweler&#39;s Forceps, Sklar 4-1/2 in</td>
			<td>Fischer Scientific</td>
			<td>50-118-4255</td>
			<td></td>
		</tr>
		<tr>
			<td>Spring Scissors - Straight/Sharp-Sharp/8mm Cutting Edge</td>
			<td>Fine Science Tools (USA) Inc.</td>
			<td>15024-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Cardiogreen</td>
			<td>Sigma</td>
			<td>I2633-25MG</td>
			<td></td>
		</tr>
		<tr>
			<td>IsosulfanBlue (Lymphazurin)&#160; 50 mg/5ml</td>
			<td>Mylan</td>
			<td>67457-220-05</td>
			<td></td>
		</tr>
	</tbody>
</table>

a murine tail lymphedema model

Lymphedema er ekstremitet hævelse forårsaget af lymfe dysfunktion. Den berørte lemmer forstørrer på grund af ophobning af væske, fedt og fibrose. Der er ingen kur mod denne sygdom. En mus hale model, der bruger en brændvidde fuld tykkelse hud excision nær bunden af halen, hvilket resulterer i hale hævelse, er blevet brugt til at studere Lymphedema. Men, denne model kan resultere i vaskulære omfatter og deraf følgende hale nekrose og tidlig hale hævelse opløsning, begrænse dens kliniske omsættelighed. Den kroniske murin hale Lymphedema model inducerer vedvarende Lymphedema over 15 uger og en pålidelig perfusion til halen. Forbedringer af den traditionelle murin hale Lymphedema model omfatter 1) præcis fuld tykkelse excision og lymfeklip ved hjælp af et kirurgisk mikroskop, 2) bekræftelse af postoperative arteriel og venøs perfusion ved hjælp af høj opløsning laser speckle, og 3) funktionel vurdering ved hjælp af indocyanin grøn nær infrarød laser lymfefiliography. Vi bruger også væv nanotransfection teknologi (TNT) for nye ikke-virale, transkutane, brændpunkt levering af genetisk last til musen hale vaskulatur.

Teknikken til musehalemodellen for vedvarende Lymphedema er vist i figur 1. Figuren udviser den relevante anatomi af musen hale model. Figur 2 viser den gradvise hævelse og vedvarende lymfedel i musehale efter Lymphedema induktion. Musen hale volumen, som beregnet af afkortet kegle ligning, toppe i uge 4 og plateauer til uge 6 efterfulgt af gradvis forbedring, der opretholdes til uge 15. Halevolumen kan bruges som en resultatvar...

Watch this Scientific Journal Video about A Murine Tail Lymphedema Model at JoVE.com

A Murine Tail Lymphedema Model

Lymphedema er ekstremitet hævelse forårsaget af lymfe dysfunktion. Vi beskriver en kronisk murin hale model af Lymphedema og den nye brug af væv nanotransfection teknologi (TNT) for genetisk last levering til halen.

En Murine Hale Lymphedema Model

Lymphedema is extremity swelling caused by lymphatic dysfunction. The affected limb enlarges because of accumulation of fluid, adipose, and fibrosis. ...

Lymphedema er en kronisk tilstand uden helbredelse. Det påvirker en tredjedel af patienter, der har haft en aksillær lymfeknude kirurgi og stråling i behandlingen af brystkræft. At have en gyldig dyremodel letter forståelsen af mekanismen og udviklingen af nye behandlingsstrategier.Musen hale model af Lymphedema er pålidelig og reproducerbar med den hurtige debut. Det udviser histologiske ændringer i overensstemmelse med human Lymphedema. Dr.Ganesh Mohan, en postdoktorgrad fra mit laboratorium.Til at begynde med skal du placere den otte uger gamle sedated mus strengt og prep halen med 70% isopropylalkohol. Brug en caliper til at måle halediameteren ved fem millimeter trin, der starter 20 millimeter fra bunden af halen. Marker en tre millimeter omskæring på halen 20 millimeter fra bunden.Under kirurgisk mikroskopisk forstørrelse, udføre en omhyggelig tre millimeter fuld tykkelse hud excision forlader alle de underliggende vaskulatur intakt. Incise den overlegne omskæring mærke først gennem dermis efterfulgt af en omskæring fuld tykkelse snit tre millimeter distal til det første snit. Lav en vinkelret fuld tykkelse lodret snit til at forbinde de to snit, derefter bruge en tandet fin pickup til at forstå en forkant og dissekere dybt inde i den avaskulære plan til dermis og overfladisk til vene adventitia med mikro saks.Indsprøjt 0,1 milliliter på 1%isosulfan blå subkutant proksimat til spidsen af halen. Identificer de to lymfekanaler vises blå på grund af injektionen støder op til den laterale hale vener. Transect lymfeknuder omhyggeligt, dissekere et plan mellem den laterale vene og lymfeknuder med lige mikrokirurgisk saks.Pass spidsen af en sakseblad mellem lymfekarret og den laterale vene og luk knivene for at transecte lymfekarret. Når du er færdig, skal du klæde halesåret med en steril klæbende klar adressering. Giv endokrine grøn 0,1 milliliter subkutant ind i den distale musehale nær spidsen.Dæmp rumlysene, og placer derefter nær infrarød laserangiografi i bufferindstilling, og udfør livebilleddannelse. Efter administration af isosulfan blå i halespidsen udviste lymfeknuderne blå farve. Lymfeknuderne blev forstyrret, mens tilstødende sideårer blev bevaret.Den progressive hævelse og vedvarende vedvarende Lymphedema i musehale efter Lymphedema induktion er vist her. Musen hale volumen som beregnet af en afkortet kegle ligning, toppede i uge fire og plateaued til uge seks efterfulgt af gradvis forbedring, der blev opretholdt indtil uge 15. Høj opløsning laser speckle kontrast billeddannelse blev gjort for at bekræfte musen hale perfusion i Lymphedema hale model viser sårede laterale vener og intakte laterale vener til vurdering af hale vaskulatur patency.Nær infrarød laser lymfangiografi demonstreret præoperative intakt lymfeknuder og ingen ICG transit ud over kirurgisk sted postoperativt, hvilket bekræfter, at hævelse var forårsaget af lymfe disfunction. Effektiviteten af genetisk last levering ved hjælp af væv nanotransfection teknologi blev demonstreret ved at levere fluorescein amidite mærket DNA til murin hale. Brug af tilpasninger til musehale Lymphedema model og ICG nær infrarød laser angiografi er en klinisk oversættes og spændende dyremodel med behandling konsekvenser.

Research

JoVE Journal

Medicine

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

En musemodel for muskeltræning ved Neuromuskulær elektrisk stimulering

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

Murine Model of Wound Healing

Musemodel af Sårheling

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

En ny musemodel af Endovaskulær aortaaneurisme Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

A Murine Model of Subarachnoid Hemorrhage

En murin model af subarachnoid blødning

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Preclinical Murine Model of Hepatic Metastases

Et præklinisk musemodel af levermetastaser

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

En murin model af Irreversible og reversibel Ensidig ureteric Obstruktion

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

Prækliniske ortotopisk Murine Model of Human Prostata Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Generating a Murine Orthotopic Metastatic Breast Cancer Model and Performing Murine Radical Mastectomy

Genererer en Murine Orthotopic metastatisk brystkræft kræft Model og udfører Murine radikale mastektomi

In vivo mouse models to assess breast cancer progression are essential for cancer research, including preclinical drug developments. However, the majority ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

Isometrisk kontraktilitet måling af musen mesenterica arterie ved hjælp af Wire blodkar

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...