Эта работа была поддержана грантами от Канадского института исследований в области здравоохранения и сердца и инсульта Фонда до н.э. и Юкон (СКК).

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+coronary+vascular+disease+in+the+heart+transplant+patient:+coronary+arteriographic+findings.">Accelerated coronary vascular disease in the heart transplant patient: coronary arteriographic findings.</a> Journal of the American College of Cardiology. 12, 334-340 (1988).</li><li>Mennander, 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=Chronic+rejection+in+rat+aortic+allografts.+An+experimental+model+for+transplant+arteriosclerosis.">Chronic rejection in rat aortic allografts. An experimental model for transplant arteriosclerosis.</a> Arterioscler Thromb. 11, 671-680 (1991).</li><li>Choy, 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=Granzymes+and+perforin+in+solid+organ+transplant+rejection.">Granzymes and perforin in solid organ transplant rejection.</a> Cell Death Differ. 17, 567-576 (2010).</li><li>Johnson, P., Carpenter, M., Hirsch, G., Lee, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recipient+cells+form+the+intimal+proliferative+lesion+in+the+rat+aortic+model+of+allograft+arteriosclerosis.">Recipient cells form the intimal proliferative lesion in the rat aortic model of allograft arteriosclerosis.</a> Am J Transplant. 2, 207-214 (2002).</li><li>Minami, E., Laflamme, M. A., Saffitz, J. E., Murry, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracardiac+progenitor+cells+repopulate+most+major+cell+types+in+the+transplanted+human+heart.">Extracardiac progenitor cells repopulate most major cell types in the transplanted human heart.</a> Circulation. 112, 2951-2958 (2005).</li><li>Yu, L., 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=AIP1+prevents+graft+arteriosclerosis+by+inhibiting+interferon-gamma-dependent+smooth+muscle+cell+proliferation+and+intimal+expansion.">AIP1 prevents graft arteriosclerosis by inhibiting interferon-gamma-dependent smooth muscle cell proliferation and intimal expansion.</a> Circ Res. 109, 418-427 (2011).</li><li>Miller, C., DeWitt, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+and+humoral+responses+to+major+and+minor+histocompatibility+antigens.">Cellular and humoral responses to major and minor histocompatibility antigens.</a> Transplant Proc. 5, 303-305 (1973).</li><li>Tsutsui, 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=Lumen+loss+in+transplant+coronary+artery+disease+is+a+biphasic+process+involving+early+intimal+thickening+and+late+constrictive+remodeling:+results+from+a+5-year+serial+intravascular+ultrasound+study.">Lumen loss in transplant coronary artery disease is a biphasic process involving early intimal thickening and late constrictive remodeling: results from a 5-year serial intravascular ultrasound study.</a> Circulation. 104, 653-657 (2001).</li><li>Rossum, A., Enns, W., Shi, P., MacEwan, G. E., Choy, 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=Bim+regulates+allogeneic+immune+responses+and+transplant+arteriosclerosis+through+effects+on+T+cell+activation+and+death.">Bim regulates allogeneic immune responses and transplant arteriosclerosis through effects on T cell activation and death.</a> Arterioscler Thromb Vasc Biol. 34, 1290-1297 (2014).</li><li>Ho, M. K., Springer, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+distribution,+structural+characterization,+and+biosynthesis+of+Mac-3,+a+macrophage+surface+glycoprotein+exhibiting+molecular+weight+heterogeneity.">Tissue distribution, structural characterization, and biosynthesis of Mac-3, a macrophage surface glycoprotein exhibiting molecular weight heterogeneity.</a> J Biol Chem. 258, 636-642 (1983).</li><li>Inoue, T., Plieth, D., Venkov, C. D., Xu, C., Neilson, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibodies+against+macrophages+that+overlap+in+specificity+with+fibroblasts.">Antibodies against macrophages that overlap in specificity with fibroblasts.</a> Kidney international. 67, 2488-2493 (2005).</li><li>Shi, 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=Immunologic+basis+of+transplant-associated+arteriosclerosis.">Immunologic basis of transplant-associated arteriosclerosis.</a> Proc Natl Acad Sci U S A. 93, 4051-4056 (1996).</li><li>Skaro, A. I., 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=CD8++T+cells+mediate+aortic+allograft+vasculopathy+by+direct+killing+and+an+interferon-gamma-dependent+indirect+pathway.">CD8+ T cells mediate aortic allograft vasculopathy by direct killing and an interferon-gamma-dependent indirect pathway.</a> Cardiovasc Res. 65, 283-291 (2005).</li><li>Tellides, 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=Interferon-gamma+elicits+arteriosclerosis+in+the+absence+of+leukocytes.">Interferon-gamma elicits arteriosclerosis in the absence of leukocytes.</a> Nature. 403, 207-211 (2000).</li><li>Wang, 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=Interferon-gamma+induces+human+vascular+smooth+muscle+cell+proliferation+and+intimal+expansion+by+phosphatidylinositol+3-kinase+dependent+mammalian+target+of+rapamycin+raptor+complex+1+activation.">Interferon-gamma induces human vascular smooth muscle cell proliferation and intimal expansion by phosphatidylinositol 3-kinase dependent mammalian target of rapamycin raptor complex 1 activation.</a> Circ Res. 101, 560-569 (2007).</li><li>Soulez, 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=The+perlecan+fragment+LG3+is+a+novel+regulator+of+obliterative+remodeling+associated+with+allograft+vascular+rejection.">The perlecan fragment LG3 is a novel regulator of obliterative remodeling associated with allograft vascular rejection.</a> Circ Res. 110, 94-104 (2012).</li><li>Choy, J. C., Kerjner, A., Wong, B. W., McManus, B. M., Granville, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perforin+mediates+endothelial+cell+death+and+resultant+transplant+vascular+disease+in+cardiac+allografts.">Perforin mediates endothelial cell death and resultant transplant vascular disease in cardiac allografts.</a> Am J Pathol. 165, 127-133 (2004).</li><li>Choy, 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=Granzyme+B+induces+endothelial+cell+apoptosis+and+contributes+to+the+development+of+transplant+vascular+disease.">Granzyme B induces endothelial cell apoptosis and contributes to the development of transplant vascular disease.</a> Am J Transplant. 5, 494-499 (2005).</li><li>Reis, E. 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=Dramatic+remodeling+of+advanced+atherosclerotic+plaques+of+the+apolipoprotein+E-deficient+mouse+in+a+novel+transplantation+model.">Dramatic remodeling of advanced atherosclerotic plaques of the apolipoprotein E-deficient mouse in a novel transplantation model.</a> Journal of vascular surgery. 34, 541-547 (2001).</li><li>Potteaux, 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=Suppressed+monocyte+recruitment+drives+macrophage+removal+from+atherosclerotic+plaques+of+Apoe-/-+mice+during+disease+regression.">Suppressed monocyte recruitment drives macrophage removal from atherosclerotic plaques of Apoe-/- mice during disease regression.</a> J Clin Invest. 121, 2025-2036 (2011).</li></ol>

Мы описали протокол для аорты промежуточным прививки у мышей, что является полезным для изучения иммунной опосредованного сосудистой неприятие и TA. Эта модель может быть использована для расследования причин ТА, а также развитие новых терапевтических стратегий. Он был использован в п...

За последние 30 с лишним лет, прогресс в иммуносупрессивных препаратов уменьшились отторжения трансплантата вследствие острого отторжения, но хроническое отторжение остается главной задачей. Основным проявлением хронического отторжения трансплантата сердце пересадки атеросклероз (ТА) 1,2. Это условие характеризуется гиперплазией интимы и вазомоторного дисфункции аллотрансплантата артерий и развивается в результате иммунологической нацеливания эндотелиальных и гладкомышечных клетках получателем иммунной системы. Таргетинга привитого сосудистой связи с признанием иностранных пептидов главного комплекса гистосовместимости (МНС) подсвечивается развития ТА исключительно в привитых артерий, щадя хозяев сосуды 3. В соответствии с этим является наблюдение, что ТА не происходит экспериментально, когда получатель является генетически идентичны донору или когда получатель не имеет Т- и В-клетки 4. Иммунная опосредованной сосудистой травмы и dysfunctioп приводит к развитию утолщением интимы и фиброза, а также аномальным накоплением липидов и белков ЕСМ в TA 5. Утолщение интимы, как правило, быть концентрическими по всей артериального дерева 4-6. Потери трансплантата и смерть обычно происходит в результате прогрессирующей ишемии вследствие окклюзии просвета артерии аллотрансплантата 4. В 1991 году Mennander др. 7 впервые аорты модель вмешательство на крысах модели TA. Несколько групп впоследствии адаптированы эту процедуру для использования в мышах. В этой модели, аллотрансплантатов аортального сегменты развиваются повреждения, которые имеют особенности, сопоставимые с ТА наблюдается в клинических трансплантаций. Это включает в себя утолщение интимы, характеризующийся накоплением гладких мышечных клеток, как и лейкоцитов получателей 7. За последние 2 десятилетия эта модель была использована для создания важное понимание механизмов повреждения сосудов, неприятия и ТП. Это может быть намиред изучить вопросы, связанные с иммунной и сосудистых реакций в ходе артериальной патологии. Выбор антигена несоответствия воздействий способность должным образом решать эти вопросы. Трансплантация по полной МНС барьеров позволяет комплексную оценку иммунного ответа, что, как известно, участвует в отторжение трансплантата. Это включает в себя прямое признание клеток CD4 и CD8 T и адресности иностранных пептид-MHC, представленный привитых клеток, полученных, косвенные CD4 (и, возможно, CD8) Т признание клеток и адресности привитых полученных аллоантигенам представленных получателя антиген-представляющих клеток, и антитело опосредованной признание аллоантигенам на поверхности клеток сосудистых 8. Тем не менее, сосудистый ответ на повреждение в полных МНС-несогласованных экспериментов может быть различным, чем это наблюдалось клинически. Джонсон и др. 9, показал, что в аорты разъединению трансплантатов, пересаженных через полное несоответствие МНС барьер, большинство изв неоинтимальная клетки реципиента происхождения, а не донорской происхождения. Это отличается от, что наблюдается в человеческих трансплантатов, где большинство внутреннюю оболочку клетки гладких мышц, донорской происхождения 9,10. Для учета этого ограничения, альтернативные экспериментальные модели, которые включают прививки по незначительных несоответствий антиген гистосовместимости были разработаны, которые вызывают сосудистые реакции, которые больше напоминают те, наблюдается в клинической трансплантации 11. В то время как эти альтернативные модели позволяют важных выводов, который можно сделать о сосудистых реакций, которые приводят в развитие ТА, иммунологических процессов, которые вызывают сосудистый отказ в Малой антиген гистосовместимости несогласованных трансплантатов не полностью заново капитулировать тех, которые происходят в клинических условиях. Например, незначительные антигены гистосовместимости признаются плохо от привитых реактивных антител 12. Учитывая вышеизложенные соображения, важно рассмотреть патологический questiна изучаются при выборе типа антигена несоответствия, используемого в модели аорты разъединения. Здесь мы опишем подробный протокол для мыши аорты разъединительных прививки. Мы описываем вмешательство прививки между полными MHC-несогласованных мышей, но тот же протокол используется для прививки через другие антиген несогласованных линиях мышей.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Name&#160;</td>
			<td>Company</td>
			<td>Catalogue</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>C57BL/6J (H-2b)</td>
			<td>Jackson Laboratories, Bar Harbour ME</td>
			<td>Strain# 000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Balb/cBYJ</td>
			<td>Jackson Laboratories, Bar Harbour ME</td>
			<td>Strain# 001026</td>
			<td></td>
		</tr>
		<tr>
			<td rowspan="2">Ketamine Hydrochloride Injection USP 100 mg/ mL</td>
			<td rowspan="2">Ketalean</td>
			<td rowspan="2">DIN 00612316</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine Injection 20 mg/mL</td>
			<td>Rompum</td>
			<td>DIN 02169592</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketoprofen Injection 100 mg/mL</td>
			<td>Anafen</td>
			<td>DIN 01938126</td>
			<td></td>
		</tr>
		<tr>
			<td>Butorphanol Tartrate&#160;injection 10 mg/mL</td>
			<td>Torbugesic</td>
			<td>DIN 008450000</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine Injection 0.3 mg/mL</td>
			<td>Reckitt Benckiser</td>
			<td>B.N. 5241</td>
			<td></td>
		</tr>
		<tr>
			<td>Atipamezole hydrochloride sterile injectable solution</td>
			<td>Antisedan</td>
			<td>DIN 02237744</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin Sodium Injection, USP, 1000 units/mL</td>
			<td>McKesson Distribution</td>
			<td>DIN 02264315</td>
			<td></td>
		</tr>
		<tr>
			<td>Tears naturale ophthalmic ointment</td>
			<td>Alcon</td>
			<td>DIN 02082519</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereomicroscope</td>
			<td>Leica</td>
			<td>M80</td>
			<td></td>
		</tr>
		<tr>
			<td>0.9% Sodium Chloride, sterile</td>
			<td>Baxter Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Lactated Ringer&#8217;s solution, sterile</td>
			<td>Baxter Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>0.9% Sodium Chloride Injection, sterile, 10 mL</td>
			<td>Baxter Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol Prep Pads</td>
			<td>Loris</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone Iodine</td>
			<td>Betadine</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Chlorohexidine Gluconate 4% w/v</td>
			<td>Germi-Stat</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Black Polyamide Monofilament</td>
			<td>AROSurgical Instruments</td>
			<td rowspan="2">T4A10Q07</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, 10-0 suture, 70 microns</td>
			<td>Corporation</td>
			<td></td>
		</tr>
		<tr>
			<td>Blue monofilament suture 5-0, P3 needle</td>
			<td>Ethicon</td>
			<td>8698G</td>
			<td></td>
		</tr>
		<tr>
			<td>1 ml Syringe</td>
			<td>BD</td>
			<td>REF 309659</td>
			<td></td>
		</tr>
		<tr>
			<td>10 ml Syringe</td>
			<td>BD</td>
			<td>REF 309604</td>
			<td></td>
		</tr>
		<tr>
			<td>1cc TB insulin syringe with 28G 1/2</td>
			<td>BD</td>
			<td>REF 309309</td>
			<td></td>
		</tr>
		<tr>
			<td>25G 7/8, hypodermic needle</td>
			<td>BD</td>
			<td>REF 305124</td>
			<td></td>
		</tr>
		<tr>
			<td>27G 1/2, hypodermic needle</td>
			<td>BD</td>
			<td>REF 305109</td>
			<td></td>
		</tr>
		<tr>
			<td>Colibri Retractor- 1.5cm spread 4cm</td>
			<td>Fine Science Tools</td>
			<td>17000-04</td>
			<td></td>
		</tr>
		<tr>
			<td>S&#38;T CAF-4 Clip applying forceps, without lock</td>
			<td>Fine Science Tools</td>
			<td>00072-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Supergrip forceps, S&#38;T</td>
			<td>Fine Science Tools</td>
			<td>00632-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical No.5 forceps</td>
			<td>Fine Science Tools</td>
			<td>11253-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Lexer Baby Scissors</td>
			<td>Fine Science Tools</td>
			<td>14078-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Adson forceps serrated</td>
			<td>Fine Science Tools</td>
			<td>11018-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Vannas-Tubingen microscissors</td>
			<td>Fine Science Tools</td>
			<td>15003-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro clamps, b-1; 3.5mm x 1mm; 7mm length</td>
			<td>Fine Science Tools</td>
			<td>00396-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe-forceps, 10cm 1x2 teeth</td>
			<td>Fine Science Tools</td>
			<td>11054-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo with lock and tungsten jaws</td>
			<td>Fine Science Tools</td>
			<td>12565-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Hot glass bead sterilizer</td>
			<td>Inotech 250</td>
			<td>IS-250 - Steri-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-woven gauzes</td>
			<td>Progene</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Tipped Applicators</td>
			<td>Puritan</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Beard Trimmer</td>
			<td>Wahl</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Sunbeam</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

mouse model of alloimmune-induced vascular rejection and transplant arteriosclerosis

Vascular rejection that leads to transplant arteriosclerosis (TA) is the leading representation of chronic heart transplant failure. In TA, the immune system of the recipient causes damage of the arterial wall and dysfunction of endothelial cells and smooth muscle cells. This triggers a pathological repair response that is characterized by intimal thickening and luminal occlusion. Understanding the mechanisms by which the immune system causes vasculature rejection and TA may inform the development of novel ways to manage graft failure. Here, we describe a mouse aortic interposition model that can be used to study the pathogenic mechanisms of vascular rejection and TA. The model involves grafting of an aortic segment from a donor animal into an allogeneic recipient. Rejection of the artery segment involves alloimmune reactions and results in arterial changes that resemble vascular rejection. The basic technical approach we describe can be used with different mouse strains and targeted interventions to answer specific questions related to vascular rejection and TA.

В этой модели, брюшной аорты у мыши Balb / cYJ вставлен в инфраренального аорты в / 6 получателя C57Bl. Это позволяет всесторонне оценить аллоиммунная ответов, которые направлены аллотрансплантатов артерии. Иммунная опосредованной травмы сосудов в этой модели инициирует сосудистых репаратив...

Watch this Scientific Journal Video about Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis at JoVE.com

Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis

We describe a protocol for aortic interposition grafting in mice. The goal of the protocol is to provide a model with which to study pathological processes and therapeutic strategies relevant to alloimmune reactions in arteries and the resultant arterial changes that contribute to organ transplant failure.

Мышь Модель аллоиммунная-индуцированной сосудистой неприятие и трансплантации атеросклероз

Vascular rejection that leads to transplant arteriosclerosis (TA) is the leading representation of chronic heart transplant failure. In TA, the immune ...

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Research

JoVE Journal

Medicine

8.4K Views.  Simon Fraser University. We describe a protocol for aortic interposition grafting in mice. The goal of the protocol is to provide a model with which to study pathological processes and therapeutic strategies relevant to alloimmune reactions in arteries and the resultant arterial changes that contribute to organ transplant failure. 

Video: Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE). In the United States (U.S.), the high morbidity and mortality rates make VTE a serious health concern 1-2. After heart disease and stroke, VTE is the third most common vascular disease 3. In the U.S. alone, there is an estimated 900,000 people affected each year, with 300,000 deaths occurring annually 3. A reliable in vivo animal model to study the mechanisms of this disease is necessary.
The advantages of using the mouse complete stasis model of inferior vena cava thrombosis are several. The mouse model allows for the administration of very small volumes of limited availability test agents, reducing costs dramatically. Most promising is the potential for mice with gene knockouts that allow specific inflammatory and coagulation factor functions to be delineated. Current molecular assays allow for the quantitation of vein wall, thrombus, whole blood, and plasma for assays. However, a major concern involving this model is the operative size constraints and the friability of the vessels. Also, due to the small IVC sample weight (mean 0.005 grams) it is necessary to increase animal numbers for accurate statistical analysis for tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent (ELISA), zymography, vein wall and thrombus cellular analysis, and whole blood and plasma assays 4-8.
The major disadvantage with the stasis model is that the lack of blood flow inhibits the maximal effect of administered systemic therapeutic agents on the thrombus and vein wall.

The mouse complete stasis model of inferior vena cava thrombosis yields quantifiable amounts of vein wall tissue and thrombus. It has proven useful for evaluating interactions between the vein wall and the occlusive thrombus and in assessing the progression from acute to chronic inflammation.

Мышь Полная модель Стасис из Нижнего тромбоз полой вены

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

Mouse Models for Graft Arteriosclerosis

Graft arteriosclerois (GA), also called allograft vasculopathy, is a pathologic lesion that develops over months to years in transplanted organs characterized by diffuse, circumferential stenosis of the entire graft vascular tree. The most critical component of GA pathogenesis is the proliferation of smooth muscle-like cells within the intima. When a human coronary artery segment is interposed into the infra-renal aortae of immunodeficient mice, the intimas could be expand in response to adoptively transferred human T cells allogeneic to the artery donor or exogenous human IFN-&gamma; in the absence of human T cells. Interposition of a mouse aorta from one strain into another mouse strain recipient is limited as a model for chronic rejection in humans because the acute cell-mediated rejection response in this mouse model completely eliminates all donor-derived vascular cells from the graft within two-three weeks. We have recently developed two new mouse models to circumvent these problems. The first model involves interposition of a vessel segment from a male mouse into a female recipient of the same inbred strain (C57BL/6J). Graft rejection in this case is directed only against minor histocompatibility antigens encoded by the Y chromosome (present in the male but not the female) and the rejection response that ensues is sufficiently indolent to preserve donor-derived smooth muscle cells for several weeks. The second model involves interposing an artery segment from a wild type C57BL/6J mouse donor into a host mouse of the same strain and gender that lacks the receptor for IFN-&gamma; followed by administration of mouse IFN-&gamma; (delivered via infection of the mouse liver with an adenoviral vector. There is no rejection in this case as both donor and recipient mice are of the same strain and gender but donor smooth muscle cells proliferate in response to the cytokine while host-derived cells, lacking receptor for this cytokine, are unresponsive. By backcrossing additional genetic changes into the vessel donor, both models can be used to assess the effect of specific genes on GA progression. Here, we describe detailed protocols for our mouse GA models.

We describe protocols for our mouse graft arteriosclerois (GA) models which involve interposition of a mouse vessel segment into a recipient of the same inbred strain. By backcrossing additional genetic changes into the vessel donor, the model can assess the effect of specific genes on GA.

Мышиные модели для взяток Атеросклероз

Graft arteriosclerois (GA),  also called allograft vasculopathy, is a pathologic lesion that develops over  months to years in transplanted organs ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Технические аспекты свища аортокавальной мышь

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Murine Heterotopic Heart Transplant Technique

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique was developed and reported in 2001 by Niimi. Described here are the techniques that have evolved over more than a decade in the hands of three surgeons (Plenter, Grazia, Pietra) in our center. These techniques are now being passed on to a younger generation of surgeons and researchers.
Based largely on the Niimi experience, the procedures used have evolved in the fine details - details which we will endeavor to relate here in such a way that others may be able to use this very useful model. Like Niimi, we have found that a video aid to learning is a priceless resource for the beginner.

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

Мышиные Гетеротопическая пересадке сердца Техника

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

Мышь трансплантации почки: Модели аллотрансплантата Отказ

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Хлорного железа, вызванной Тромбоз мышь Модель на сонной артерии и брыжейки судна

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The simple murine AVF model was combined with intraluminal administration of drug solution to the venous endothelium at the same time as fistula creation. Technical aspects of this model are discussed. Under general anesthesia, an abdominal incision is made and the aorta and inferior vena cava (IVC) are exposed. The infra-renal aorta and IVC are dissected for clamping. After proximal and distal clamping, the puncture site is exposed and a 25 G needle is used to puncture both walls of the aorta and into the IVC. Immediately after the puncture, a reporter gene-expressing viral vector was infused in the IVC via the same needle, followed by 15 min of incubation. The intraluminal administration method enabled more robust viral gene delivery to the venous endothelium compared to administration by the external route. This novel method of delivery will facilitate studies that explore the role of the endothelium in AVF maturation and enable intraluminal drug delivery at the time of surgical operation.

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

Внутрипросветный доставки лекарств к мыши артериовенозной фистулы эндотелия

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

Жила интерпозиция Модель: A Подходит модель для изучения шунтирование проходимость

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

Использование метода рукав в мышиной модели аортальной трансплантации - обучающее видео

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of aortic disease exist, in vivo imaging of the vasculature has been limited. In recent years, micro-computerized tomography (micro-CT) has emerged as a preferred modality for imaging both large and small vessels both in vivo and ex vivo. In conjunction with a method of vascular casting, we have successfully used micro-CT to characterize the frequency and distribution of aortic pathology in &#946;-aminopropionitrile-treated C57/Bl6 mice. Technical limitations of this method include variations in the quality of the perfusion introduced by poor animal preparation, the application of proper methodologies for vessel size quantification, and the non-survivability of this procedure. This article details a methodology for the intravascular perfusion of a lead-based radiopaque silicone rubber for the quantitative characterization of aortopathy in a mouse model of aneurysm and dissection. In addition to visualizing aortic pathology, this method may be used for examining other vascular beds in vivo or vascular beds removed post-mortem.

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &amp;#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

This article describes a detailed methodology of using a radiopaque lead-based silicone rubber to perfuse the murine vasculature for aortic diameter quantification in a mouse model of aortic aneurysm and dissection.

Количественный анализ микро CT Aortopathy в мышиной модели β-aminopropionitrile индуцированной аневризмы аорты и рассечение

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...