This work was supported by the American Heart Association Scientist Development Grant and the Foundation for Anesthesia Education and Research (MLJ). We would like to thank Talaignair N. Venkatraman PhD for his assistance with magnetic resonance imaging.

Etik Beyanı: Bu protokol, Duke Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onaylanmış ve hayvanların etik kullanımı için tüm kuralları takip edilmiştir. Ekipman 1. Hazırlanması <ol><li> Ameliyat öncesi cerrahi aletler otoklavlayın. </li><li> % 70 etanol ile stereotaktik aparatı dezenfekte edin. </li><li> Su banyosu açın ve 42 ° C&#39;de su sıcaklığı tutmak </li><li> 0.4 ul başına 0.075 U &#39;lik bir konsantrasyonda, normal tuzlu su içinde Type IV-S klostridial kolajenaz çözülür. </li></ol> 2.. Kolajenaz Enjeksiyon Model <ol><li> Fare tartılır. </li><li> % 30 O 2 /% 70 N 2% 5 izofluran ile indüksiyon odasında fare anestezisi. Yeterli anestezi fare solunum saniyede 1 yavaşladı yaklaşık 2 dakika sonra bildirilir. </li><li> 30 mm 20 G intravenöz kateter ile trakea entübe. </li><li> Bir kateteri geçkemirgen vantilatör ve mekanik olarak% 30% 1.6 izofluran ile akciğerlere havalandırmak O 2/70 0.75 ml &#39;lik bir teslim tidal hacmi ile dakikada 105 nefes&#39; lik bir oranda% 2 .. </li><li> Bir elektronik tıraş makinesi ile kafa derisi tıraş. Fare uyutuldu ve sonda sonra, traş etme için başka bir çalışma istasyonuna taşımak ve daha sonra cerrahi tezgah döndü. </li><li> Stereotaktik çerçeve içinde başını sağlamlaştırmak, ve referans noktaları olarak koronal ve sagital hem sütür ile baş seviyesi. </li><li> Gözleri oftalmik merhem sürün. </li><li> Bir rektal ısı probu yerleştirin. Su yatağı dolaşan bir alt gövdeyi kullanarak 37.0 ± 0.2 ° C rektal sıcaklığı korumak. </li><li> Wipe betadin cerrahi alan% 70 etanol ile takip ve 3 kez tekrarlayın. </li><li> 1 cm orta hat kafa derisi kesi yapmak ve bregma maruz steril pamuk uçlu bir aplikatör ile yanal periosta silin. </li><li> Matkap çapı 1 mm burr hole 2,2 mm lat bıraktısu ile soğutulan bir matkap ile bregmaya eral. </li><li> Kolajenaz şişe 5 çevirin, daha sonra 0.5 ul kolajenaz çözeltisi (stereotaksik çerçeveye bağlı) 25 G iğne ile 5 kez (Son yıkamadan sonra, şırınga içinde kolajenaz çözeltisi 0.5 ul bırak), bir 0.5 ul şırınga yıkayın. </li><li> Çapak delik ile iğne ucu hizalamak şırınga 0.1 ul sınırdışı ve atmak için ustura ile iğne konik silin. </li><li> Bir mikromanipülatör kullanarak, kortekse derin iğne 3 mm ilerlemek ve 30 saniye boyunca hareketsiz bırakın. </li><li> 90 saniye boyunca 0.4 ul enjekte edilir. </li><li> % 1 izofluran azaltmak ve 5 dakika boyunca iğne hareketsiz bırakın. </li><li> Yavaş yavaş iğneyi. </li><li> % 0.25 bupivakain subkutan 2 damla ve cilt sütür 1 - uygulayın. </li><li> Izofluran buharlaştırıcı kapatın ve stereotaktik çerçeve fareyi çıkarın. </li><li> Fare sonraki ekstübasyon ile spontan ventilasyon kurtarmak için izin verin. </li><li> Temiz bir kafes fare dönün ve ücretsiz izinYiyecek ve su. </li></ol> 3.. Otolog Kan Enjeksiyon Modeli <ol><li> Kollajenaz enjeksiyon modeli için 2.11 - 2.1 adımları izleyin. </li><li> Bir 30 G 50 ul şırınga içine steril normal tuzlu su, 50 ul çizin. </li><li> 70 cm PE10 boru ile mikrolitre şırınga bağlayın. </li><li> PE10 tüp tamamen de-hava tüpü içine mikrolitre şırınga tüm normal tuzlu kovun. </li><li> Daha sonra tuzlu su ve işlemler sırasında kan karışımı önlemek için PE10 boru mikrolitre şırınga düzeneğinin distal açıklık bir hava kabarcığı yapmak için dışarı 1 mm mikrolitre şırınga pistonunu çıkarın. </li><li> % 70 etanol ile fare distal santral kuyruk arter bölgeyi silin, ve kuyruk ucuna 1 cm 0.5 de bir ustura ile arter kesti. </li><li> PE10 boru mikrolitre şırınga düzeneğinin kesilmiş kuyruk kan 40 ul toplayın. Not: heparin, iğne, boru, ya da fare kullanılmaz. </li><li> Inje üzerine mikrolitre şırınga takınction pompası. </li><li> PE10 tüp sonuna bir 27 G iğne metal kanül kısmı bağlayın ve stereotaktik çerçeve üzerinde bir micromanipulator iğne sabitleyin. </li><li> 27 G iğne üzerinden kan 2 ul sınırdışı ve atmak için ustura ile iğne konik silin. </li><li> Çapak delik ile iğne ucu hizalayın ve kortekse derin iğne 3 mm yerleştirin. </li><li> Dakika başına 2 ul &#39;lik bir oranda otolog kan 35 ul enjekte edilir. </li><li> % 1 izofluran azalma ve 10 dakika boyunca iğne hareketsiz bırakın. </li><li> 30 saniye içinde iğneyi. </li><li> % 0.25 bupivakain subkutan 2 damla ve cilt sütür 1 - uygulayın. </li><li> Izofluran buharlaştırıcı kapatın ve stereotaktik çerçeve fareyi çıkarın. </li><li> Fare sonraki ekstübasyon ile spontan ventilasyon kurtarmak için izin verin. </li><li> Temiz bir kafes fare dönün ve gıda ve su ücretsiz erişim sağlar. </li></ol> 4. Sham Operasyonu <ol><li> Kollajenaz injectio için aynı prosedürleri takipiğne yerleştirildikten sonra enjeksiyon olmadan hariç n modeli. </li></ol> 5. Post-cerrahi Bakımı <ol><li> Hayvanın boynun arkasındaki cerrahi prosedürün akşam normal tuzlu su deri altına, 0.5 ml enjekte edilir. </li><li> Kafesin zeminine yerleştirilen küçük plastik bardak, su ve jel gıda ile yumuşatılmış yiyecek sağlamak. 7 gün boyunca günlük yiyecek değiştirin. </li><li> 7 gün boyunca günlük ağırlık kaybı, yara iyileşmesi ve rahatsızlık belirtileri kontrol edin. </li><li> 7 günden daha fazla geri kazanımı aralıkları gerekmektedir, gerekirse, ameliyat dikiş ipliği kaldırma, (% 30, O 2/70, N% 2 yaklaşık olarak% 1 izofluran) açık inhale anestezi altında gerçekleştirilebilir. </li></ol>

<ol>
	<li>Asch, C. 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=Incidence,+case+fatality,+and+functional+outcome+of+intracerebral+haemorrhage+over+time,+according+to+age,+sex,+and+ethnic+origin:+a+systematic+review+and+meta-analysis.">Incidence, case fatality, and functional outcome of intracerebral haemorrhage over time, according to age, sex, and ethnic origin: a systematic review and meta-analysis.</a> Lancet Neurology. 9, 167-176 (2010).</li><li>Qureshi, A. I., Mendelow, A. D., Hanley, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracerebral+haemorrhage.">Intracerebral haemorrhage.</a> Lancet. 373, 1632-1644 (2009).</li><li>Participants, N. I. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Priorities+for+clinical+research+in+intracerebral+hemorrhage:+report+from+a+National+Institute+of+Neurological+Disorders+and+Stroke+workshop.">Priorities for clinical research in intracerebral hemorrhage: report from a National Institute of Neurological Disorders and Stroke workshop.</a> Stroke. 36, (2005).</li><li>Morgenstern, L. B., 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=Guidelines+for+the+management+of+spontaneous+intracerebral+hemorrhage:+a+guideline+for+healthcare+professionals+from+the+American+Heart+Association/American+Stroke+Association.">Guidelines for the management of spontaneous intracerebral hemorrhage: a guideline for healthcare professionals from the American Heart Association/American Stroke Association.</a> Stroke. 41, 2108-2129 (2010).</li><li>James, M. L., Warner, D. S., Laskowitz, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+models+of+intracerebral+hemorrhage:+a+translational+perspective.">Preclinical models of intracerebral hemorrhage: a translational perspective.</a> Neurocrit Care. 9, 139-152 (2008).</li><li>Winkler, D. 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=Spontaneous+hemorrhagic+stroke+in+a+mouse+model+of+cerebral+amyloid+angiopathy.">Spontaneous hemorrhagic stroke in a mouse model of cerebral amyloid angiopathy.</a> J Neurosci. 21, 1619-1627 (2001).</li><li>Sinar, E. J., Mendelow, A. D., Graham, D. I., Teasdale, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+intracerebral+hemorrhage:+effects+of+a+temporary+mass+lesion.">Experimental intracerebral hemorrhage: effects of a temporary mass lesion.</a> J Neurosurg. 66, 568-576 (1987).</li><li>Mendelow, A. D., Bullock, R., Teasdale, G. M., Graham, D. I., McCulloch, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracranial+haemorrhage+induced+at+arterial+pressure+in+the+rat.+Part+2:+Short+term+changes+in+local+cerebral+blood+flow+measured+by+autoradiography.">Intracranial haemorrhage induced at arterial pressure in the rat. Part 2: Short term changes in local cerebral blood flow measured by autoradiography.</a> Neurol Res. 6, 189-193 (1984).</li><li>Bullock, R., Mendelow, A. D., Teasdale, G. M., Graham, D. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracranial+haemorrhage+induced+at+arterial+pressure+in+the+rat.+Part+1:+Description+of+technique,+ICP+changes+and+neuropathological+findings.">Intracranial haemorrhage induced at arterial pressure in the rat. Part 1: Description of technique, ICP changes and neuropathological findings.</a> Neurol Res. 6, 184-188 (1984).</li><li>Sang, Y. H., Su, H. X., Wu, W. T., So, K. F., Cheung, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elevated+blood+pressure+aggravates+intracerebral+hemorrhage-induced+brain+injury.">Elevated blood pressure aggravates intracerebral hemorrhage-induced brain injury.</a> J Neurotrauma. 28, 2523-2534 (2011).</li><li>Krafft, P. 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=Modeling+intracerebral+hemorrhage+in+mice:+injection+of+autologous+blood+or+bacterial+collagenase.">Modeling intracerebral hemorrhage in mice: injection of autologous blood or bacterial collagenase.</a> J Vis Exp. , (2012).</li><li>Sansing, L. 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=Autologous+blood+injection+to+model+spontaneous+intracerebral+hemorrhage+in+mice.">Autologous blood injection to model spontaneous intracerebral hemorrhage in mice.</a> J Vis Exp. , (2011).</li><li>Rosenberg, G. A., Mun-Bryce, S., Wesley, M., Kornfeld, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collagenase-induced+intracerebral+hemorrhage+in+rats.">Collagenase-induced intracerebral hemorrhage in rats.</a> Stroke. 21, 801-807 (1990).</li><li>Nath, F. P., Jenkins, A., Mendelow, A. D., Graham, D. I., Teasdale, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+hemodynamic+changes+in+experimental+intracerebral+hemorrhage.">Early hemodynamic changes in experimental intracerebral hemorrhage.</a> J Neurosurg. 65, 697-703 (1986).</li><li>Anderson, C. 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=Rapid+blood-pressure+lowering+in+patients+with+acute+intracerebral+hemorrhage.">Rapid blood-pressure lowering in patients with acute intracerebral hemorrhage.</a> N Engl J Med. 368, 2355-2365 (2013).</li><li>Clark, W., Gunion-Rinker, L., Lessov, N., Hazel, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Citicoline+treatment+for+experimental+intracerebral+hemorrhage+in+mice.">Citicoline treatment for experimental intracerebral hemorrhage in mice.</a> Stroke. 29, 2136-2140 (1998).</li><li>Mayer, S. 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=Efficacy+and+safety+of+recombinant+activated+factor+VII+for+acute+intracerebral+hemorrhage.">Efficacy and safety of recombinant activated factor VII for acute intracerebral hemorrhage.</a> N Engl J Med. 358, 2127-2137 (2008).</li><li>Mendelow, A. 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=Early+surgery+versus+initial+conservative+treatment+in+patients+with+spontaneous+supratentorial+lobar+intracerebral+haematomas+(STICH+II):+a+randomised+trial.">Early surgery versus initial conservative treatment in patients with spontaneous supratentorial lobar intracerebral haematomas (STICH II): a randomised trial.</a> Lancet. , (2013).</li><li>James, M. L., Blessing, R., Bennett, E., Laskowitz, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apolipoprotein+E+modifies+neurological+outcome+by+affecting+cerebral+edema+but+not+hematoma+size+after+intracerebral+hemorrhage+in+humans.">Apolipoprotein E modifies neurological outcome by affecting cerebral edema but not hematoma size after intracerebral hemorrhage in humans.</a> J Stroke Cerebrovasc Dis. 18, 144-149 (2009).</li><li>James, M. L., Blessing, R., Phillips-Bute, B. G., Bennett, E., Laskowitz, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=S100B+and+brain+natriuretic+peptide+predict+functional+neurological+outcome+after+intracerebral+haemorrhage.">S100B and brain natriuretic peptide predict functional neurological outcome after intracerebral haemorrhage.</a> Biomarkers. 14, 388-394 (2009).</li><li>James, M. L., Sullivan, P. M., Lascola, C. D., Vitek, M. P., Laskowitz, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacogenomic+effects+of+apolipoprotein+e+on+intracerebral+hemorrhage.">Pharmacogenomic effects of apolipoprotein e on intracerebral hemorrhage.</a> Stroke. 40, 632-639 (2009).</li><li>James, M. 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=Brain+natriuretic+peptide+improves+long-term+functional+recovery+after+acute+CNS+injury+in+mice.">Brain natriuretic peptide improves long-term functional recovery after acute CNS injury in mice.</a> J Neurotrauma. 27, 217-228 (2010).</li><li>Indraswari, F., 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=Statins+improve+outcome+in+murine+models+of+intracranial+hemorrhage+and+traumatic+brain+injury:+a+translational+approach.">Statins improve outcome in murine models of intracranial hemorrhage and traumatic brain injury: a translational approach.</a> J Neurotrauma. 29, 1388-1400 (2012).</li><li>Laskowitz, D. 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=The+apoE-mimetic+peptide,+COG1410,+improves+functional+recovery+in+a+murine+model+of+intracerebral+hemorrhage.">The apoE-mimetic peptide, COG1410, improves functional recovery in a murine model of intracerebral hemorrhage.</a> Neurocrit Care. 16, 316-326 (2012).</li><li>Lei, B., 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=Interaction+between+sex+and+apolipoprotein+E+genetic+background+in+a+murine+model+of+intracerebral+hemorrhage.">Interaction between sex and apolipoprotein E genetic background in a murine model of intracerebral hemorrhage.</a> Translational Stroke Research. 3, (2012).</li><li>Lekic, 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=Evaluation+of+the+hematoma+consequences,+neurobehavioral+profiles,+and+histopathology+in+a+rat+model+of+pontine+hemorrhage.">Evaluation of the hematoma consequences, neurobehavioral profiles, and histopathology in a rat model of pontine hemorrhage.</a> J Neurosurg. 118, 465-477 (2013).</li><li>Nakamura, 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=Intracerebral+hemorrhage+in+mice:+model+characterization+and+application+for+genetically+modified+mice.">Intracerebral hemorrhage in mice: model characterization and application for genetically modified mice.</a> J Cereb Blood Flow Metab. 24, 487-494 (2004).</li><li>Yang, 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=Statins+Protect+the+Blood+Brain+Barrier+Acutely+after+Experimental+Intracerebral+Hemorrhage.">Statins Protect the Blood Brain Barrier Acutely after Experimental Intracerebral Hemorrhage.</a> J Behav Brain Sci. 3, 100-106 (2013).</li><li>Rynkowski, M. 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=A+mouse+model+of+intracerebral+hemorrhage+using+autologous+blood+infusion.">A mouse model of intracerebral hemorrhage using autologous blood infusion.</a> Nature Protocols. 3, 122-128 (2008).</li><li>Wang, J., Fields, J., Dore, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+an+improved+preclinical+mouse+model+of+intracerebral+hemorrhage+using+double+infusion+of+autologous+whole+blood.">The development of an improved preclinical mouse model of intracerebral hemorrhage using double infusion of autologous whole blood.</a> Brain Research. 1222, 214-221 (2008).</li><li>MacLellan, C. 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=Intracerebral+hemorrhage+models+in+rat:+comparing+collagenase+to+blood+infusion.">Intracerebral hemorrhage models in rat: comparing collagenase to blood infusion.</a> J Cereb Blood Flow Metab. 28, 516-525 (2008).</li></ol>

Lei, Sheng, Wang, Lascola, Warner, and Laskowitz have no conflicts of interest to declare. James received grant funding by American Heart Association, National Institutes of Health, and Cephalogics.

Therapeutics 15-18 umut verici için klinik öncesi araştırma ve sonuçta büyük klinik çalışmalar gelişmekte olmasına rağmen, orada ICH içinde sonuçlarını iyileştirdiği gösterilmiş hiçbir farmakolojik müdahaleler ve bakım büyük ölçüde destekleyici kalır. Muhtemel tedavilerin listeleri gibi Transkriptomik ve proteomik çalışma gibi yüksek kapasiteli teknolojileri ile üretilebilir. Bu teknolojiler, ileri potansiyel terapötik hedeflerin bilgimizi ilerlemeye devam ediyor ve umut ve...

Intraserebral kanama (İSK) etkilenen hastaların yaklaşık% 40-50 30 gün içinde ölme 1 ile serebrovasküler hastalık nispeten sık görülen şeklidir. Ne yazık ki, küçük bir iyileşme son 20 yıl boyunca 2 ölüm oranında yapılmıştır. Ulusal Sağlık Enstitüleri 3 ve Amerikan Kalp Derneği 4 kılavuzların raporları patofizyolojisi anlayış genişletmek ve yeni tedavi yaklaşımları için hedeflerini geliştirmeye ICH klinik olarak ilgili modellerinin geliştirilmesinin önemini vurguladı. Çeşitli modeller, insan ICH 5 taklit var. ICH patofizyoloji yetişkinlerin anlama olarak, çeşitli modeller hastalığın farklı yönlerini incelemek için kullanılabilir, belirgin hale gelmiştir. Daha önce kullanılan modeller kemirgen amiloid anjiopati 6, intraparenkimal mikrobalon ekleme ve enflasyon 7 ve doğrudan arteriyel kan bulunmaktadırinfiltrasyon 8,9. Amiloid anjiyopati ikinci Lobar kanama transgenik farelerin kullanılması ile model ve ayrı bir alt tipi ICH temsil edilmiştir. Mikrobalonların modelleri hematom akut kütle etkisinin taklit eden ancak, kan varlığı için beynin hücresel yanıtı yakalamak için başarısız. Son olarak, doğrudan arteriyel kan sızması femoral arterden arteriyel baskılara beyin tabi. Dolayısıyla, bu model taklit arter basınçları ve kan varlığı ancak küçük kan damarı yırtılması gelen mikrovasküler yaralanma beyin tabi değildir. Bundan başka, bu model, doğal olarak yüksek bir değişkenlik vardır. Onlar yaş olarak İlginçtir, spontan hipertansif sıçanlar 10 spontan ICH gelişir. ICH geliştirme sonra bu hayvanların çalışma İSH&#39;lı için insanları yatkınlık önemli Komorbiditelerin birinin varlığında hastalığı taklit edebilir. Klostridial kolajenaz 11 veya a instrastiatal enjeksiyonu bu model mevcut olmakla birlikte, enjeksiyon intrastriatalutologous tüm kan 12, şu anda en yaygın iki model ICH klinik öncesi araştırmalarda kullanılır. ICH modeli seçimi türlerin seçimi ve hematom oluşumunu sağlama yöntemi de dahil olmak deneysel sorusunun objektif dayalı yapılmalıdır. Örneğin, domuz farelere göre daha büyük bir beyaz madde beyin hacmi ile büyük hayvanlar olmalarıdır. Böylece, domuz modeller İSH&#39;lı aşağıdaki beyaz cevher patofizyolojisini incelemek için uygundur. Aksine, kemirgen beyin büyük ölçüde gri madde vardır, ancak transgenik sistemleri İSH&#39;lı yaralanma ve iyileşme moleküler mekanizmalarını değerlendirmek için kemirgenler yararlı olun. Her model dikkatli bir deney öncesinde dikkat edilmesi gereken, kendine özgü güçlü ve zayıf yönlerini (Tablo 1) vardır. Aşağıdaki protokoller farelerde otolog kan ve kolajenaz enjeksiyon modellerini göstermektedir. Bu modeller her biri aslında sıçanlarda geliştirilen modellerden çevrilmiştir13,14 ve İSH&#39;lı hücre ölümü ile ilişkili moleküler mekanizmalarını keşfetmek için yaygın olarak kullanılabilir transgenik teknolojinin kullanımına izin. Hem insan İSH&#39;lı belirgin farklı bir yaralanma mekanizmalarını ifade eder, hem de davranışsal ve histolojik önlemler açısından belirgin farklı beklenen sonuç var. Böylece, bazı hipotezler diğer üzerinden bir modele kendilerini borç olabilir, ancak birçok fikirler hem de model doğrulama gerekebilir. Kollajenaz-ve otolog kan enjeksiyonu intraserebral kanama modellerinin özellikleri Tablo 1.. Karşılaştırılması. <table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td style="width:197px;"></td><td style="width:80px;"> Kolajenaz Enjeksiyon </td><td style="width:66px;"> Kan Enjeksiyon </td></tr><tr><td style="width:197px;"> Kullanım kolaylığı </td><td style="width:80px;"> + + + </td><td style="width:66px;"> + + </td></tr><tr&gt; <td style="width:197px;"> Tekrarlanabilirlik </td><td style="width:80px;"> + + </td><td style="width:66px;"> + + </td></tr><tr><td style="width:197px;"> Kanama Boyutu Kontrolü </td><td style="width:80px;"> + + </td><td style="width:66px;"> + + + </td></tr><tr><td style="width:197px;"> Kan Reflü </td><td style="width:80px;"> + </td><td style="width:66px;"> + + </td></tr><tr><td style="width:197px;"> İnsan Hastalığı simüle </td><td style="width:80px;"> + </td><td style="width:66px;"> - </td></tr><tr><td style="width:197px;"> Basitlik </td><td style="width:80px;"> + + </td><td style="width:66px;"> + + </td></tr><tr><td style="width:197px;"> Çoklu Türler kullanın </td><td style="width:80px;"> + + </td><td style="width:66px;"> + + </td></tr></tbody></table>

<table border="0" cellpadding="0" cellspacing="0" width="416">
	<tbody>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">Name</td>
			<td style="width:88px;">Company</td>
			<td style="width:88px;">Catalog Number</td>
			<td style="width:88px;">Comments</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Stereotactic frame</td>
			<td style="width:88px;">Stoelting Co.</td>
			<td style="width:88px;">51603</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:152px;">Probe holder with corner clamp</td>
			<td style="width:88px;">Stoelting Co.</td>
			<td style="width:88px;">51631</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">Mini grinder</td>
			<td style="width:88px;">Power Glide</td>
			<td style="width:88px;">Model 60100002</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">0.5 &#181;l Hamilton syringe</td>
			<td style="width:88px;">Hamilton Co.</td>
			<td style="width:88px;">86259</td>
			<td style="width:88px;">25 gauge needle</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">50 &#181;l Hamilton syringe</td>
			<td style="width:88px;">Hamilton Co</td>
			<td style="width:88px;">7637-01</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:152px;">26G Hamilton needle</td>
			<td style="width:88px;">Hamilton Co</td>
			<td style="width:88px;">7804-03</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Syringe pump</td>
			<td style="width:88px;">KD Scientific</td>
			<td style="width:88px;">Model 100</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:152px;">Heat therapy water pump</td>
			<td style="width:88px;">Gaymar Industries, Inc.</td>
			<td style="width:88px;">Model# TP650</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">Circulating waterbed</td>
			<td style="width:88px;">CMS Tool &#38; Die, Inc.</td>
			<td style="width:88px;"></td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">Rodent ventilator</td>
			<td style="width:88px;">Harvard Apparatus</td>
			<td style="width:88px;">Model 683</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Isoflurane vaporizer</td>
			<td style="width:88px;">Drager</td>
			<td style="width:88px;">Vapor 19.1</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:152px;">Air flowmeter</td>
			<td style="width:88px;">Cole Parmer</td>
			<td style="width:88px;">Model PMR1-010295</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Induction chamber</td>
			<td style="width:88px;"></td>
			<td style="width:88px;"></td>
			<td style="width:88px;">Self made</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Otoscope</td>
			<td style="width:88px;">Welch Allyn</td>
			<td style="width:88px;">22820</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:152px;">intravenous catheter</td>
			<td style="width:88px;">Becton-Dickinson</td>
			<td style="width:88px;">381534</td>
			<td style="width:88px;">20-gauge, 1.16 inch Insyte-W</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:152px;">Isoflurane</td>
			<td style="width:88px;">Baxter Healthcare Corporation</td>
			<td style="width:88px;">NDC10019-360-69</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Collagenase Type IV-S</td>
			<td style="width:88px;">Sigma</td>
			<td style="width:88px;">C1889</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">Polyethylene tubing PE20</td>
			<td style="width:88px;">Becton-Dickinson</td>
			<td style="width:88px;">427406</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">Polyethylene tubing PE10</td>
			<td style="width:88px;">Becton-Dickinson</td>
			<td style="width:88px;">427401</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">30G 1 inch needle</td>
			<td style="width:88px;">Becton-Dickinson</td>
			<td style="width:88px;">305128</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:152px;">27G 1 1/4 inch needle</td>
			<td style="width:88px;">Becton-Dickinson</td>
			<td style="width:88px;">305136</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Surgical scissors</td>
			<td style="width:88px;">Miltex</td>
			<td style="width:88px;">21-539</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Forceps</td>
			<td style="width:88px;">Miltex</td>
			<td style="width:88px;">17-307</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Needle holder</td>
			<td style="width:88px;">Boboz</td>
			<td style="width:88px;">RS-7840</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Monofilament suture</td>
			<td style="width:88px;">Ethicon</td>
			<td style="width:88px;">8698</td>
			<td style="width:88px;">Size 5-0</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:152px;">Indicating controller</td>
			<td style="width:88px;">YSI</td>
			<td style="width:88px;">73ATD</td>
			<td style="width:88px;"></td>
		</tr>
	</tbody>
</table>

intrastriatal injection of autologous blood or clostridial collagenase as murine models of intracerebral hemorrhage

Intraserebral kanama (İSK) serebrovasküler hastalık yaygın bir şeklidir ve önemli morbidite ve mortalite ile ilişkilidir. Hemostaz ve pıhtı çıkarılması hedefleniyor büyük klinik çalışmaların etkili tedavi ve başarısızlık eksikliği ICH daha mekanizması odaklı soruşturma için ihtiyaç göstermektedir. Bu araştırma, klinik öncesi modellerde tarafından sağlanan çerçeve aracılığıyla gerçekleştirilebilir. Popüler kullanımı iki kemirgen modelleri, otolog kan veya klostridial kolajenaz ya intrastriatal (bazal ganglia) enjeksiyon yer alır. Yana, her model İSH&#39;lı ilgili belirgin farklı patofizyolojik özellikleri temsil eder, belli bir modelin kullanımı öğretilmektedir hastalığın hangi yönü dayalı seçilebilir. Örneğin, otolog kan enjeksiyonu en doğru intraparenkimal kan varlığı için beynin yanıtı temsil eder, ve en yakın lobar kanama çoğaltabilir. Clostridial kollajenaz enjeksiyon en doğru s temsilalışveriş merkezi damar rüptürü ve derin kanamalar hematom evrimi karakteristik. Böylece farklı hematom oluşumu, nöroenflamatuar tepki, serebral ödem gelişmesi ve nörodavranışsal sonuçları her model sonuçları. Sözde bir terapötik müdahale Sağlamlık en iyi iki model kullanılarak değerlendirilebilir. Bu protokol, her iki model, yaralanma hemen ameliyat sonrası gösteri ve erken post-operatif bakım teknikleri kullanılarak Vaginal indüksiyon gösterilmiştir. Her iki model de tekrarlanabilir yaralanmaları, hematom hacimleri ve nörodavranışsal kayıpla sonuçlanabilir. Çünkü insan ICH heterojenite, birden fazla preklinik modeller iyice patofizyolojik mekanizmalarını keşfetmek ve potansiyel tedavi stratejileri test etmek için gereklidir.

Hematom genişleme (Şekil 2) oluştuğunda, kolagenaz enjeksiyondan sonra 4 saat - Due hematom oluşumu (Şekil 1) farklılıklara, ipsilateral dönüm hemen otolog kan enjekte edilen farelerin için uyanmak sonra ve 2 içinde gösterilmiştir. Ipsilateral torna yokluğu önemli yaralanma olmaması için endişe yükseltmek gerekir. İlk sonrası yaralanma gününde, her iki modelde de fareler önemli nörolojik defisit (Şekil 3) göstermelidir. Enjeksiyonundan 24 saa...

Watch this Scientific Journal Video about Intrastriatal Injection of Autologous Blood or Clostridial Collagenase as Murine Models of Intracerebral Hemorrhage at JoVE.com

Intrastriatal Injection of Autologous Blood or Clostridial Collagenase as Murine Models of Intracerebral Hemorrhage

Preclinical models of intracerebral hemorrhage are utilized to mimic certain aspects of clinical disease. Thus, mechanisms of injury and potential therapeutic strategies may be explored. In this protocol, two models of intracerebral hemorrhage are described, intrastriatal (basal ganglia) injections of autologous blood or collagenase.

İntraserebral Kanama Fare Modellerinden olarak Otolog Blood veya Klostridyal Kollajenazının intrastriatal Enjeksiyon

Intracerebral hemorrhage (ICH) is a common form of cerebrovascular disease and is associated with significant morbidity and mortality. Lack of effective ...

intrastriatal-injection-autologous-blood-or-clostridial-collagenase

Research

JoVE Journal

Medicine

14.6K Görüntüleme.  Duke University. Preclinical models of intracerebral hemorrhage are utilized to mimic certain aspects of clinical disease. Thus, mechanisms of injury and potential therapeutic strategies may be explored. In this protocol, two models of intracerebral hemorrhage are described, intrastriatal (basal ganglia) injections of autologous blood or collagenase. 

Video: Intrastriatal Injection of Autologous Blood or Clostridial Collagenase as Murine Models of Intracerebral Hemorrhage

Fixed Volume or Fixed Pressure: A Murine Model of Hemorrhagic Shock

It is common knowledge that severe blood loss and traumatic injury can lead to a cascade of detrimental signaling events often resulting in mortality. 1, 2, 3, 4, 5 These signaling events can also lead to sepsis and/or multiple organ dysfunction (MOD). 6, 7, 8, 9 It is critical then to investigate the causes of suppressed immune function and detrimental signaling cascades in order to develop more effective ways to help patients who suffer from traumatic injuries. 10 This fixed pressure Hemorrhagic Shock (HS) procedure, although technically challenging, is an excellent resource for investigation of these pathophysiologic conditions. 11, 12, 13 Advances in the assessment of biological systems, i.e. Systems Biology have enabled the scientific community to further understand complex physiologic networks and cellular communication patterns. 14 Hemorrhagic Shock has proven to be a vital tool for unveiling these cellular communication patterns as they relate to immune function. 15, 16, 17, 18 This procedure can be mastered! This procedure can also be used as either a fixed volume or fixed pressure approach. We adapted this technique in the murine model to enhance research in innate and adaptive immune function. 19, 20, 21 Due to their small size HS in mice presents unique challenges. However due to the many available mouse strains, this species represents an unparalleled resource for the study of the biologic responses. The HS model is an important model for studying cellular communication patterns and the responses of systems such as hormonal and inflammatory mediator systems, and danger signals, i.e. DAMP and PAMP upregulation as it elicits distinct responses that differ from other forms of shock. 22, 23, 24, 25 The development of transgenic murine strains and the induction of biologic agents to inhibit specific signaling have presented valuable opportunities to further elucidate our understanding of the up and down regulation of signal transduction after severe blood loss, i.e. HS and trauma 26, 27, 28, 29, 30. 
There are numerous resuscitation methods (R) in association with HS and trauma. 31, 32, 33, 34 A fixed volume resuscitation method of solely lactated ringer solution (LR), equal to three times the shed blood volume, is used in this model to study endogenous mechanisms such as remote organ injury and systemic inflammation. 35, 36, 38 This method of resuscitation is proven to be effective in evaluating the effects of HS and trauma 38, 39.

The Hemorrhagic Shock model has been a reliable and reproducible resource facilitating the identification and understanding of signaling cascades associated with inflammation and end-organ damage after trauma. This article provides a step-by-step description of surgical and mechanical aspects associated with the Hemorrhagic Shock experimental procedure in mice.

Sabit Hacim ve Sabit Basınç: Hemorajik Şok murin Modeli

It is common knowledge that severe blood loss and traumatic injury can lead to a cascade of detrimental signaling events often resulting in mortality. 1, ...

Tıp

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo. With the advent of genetic modifications to the whole organism or the myocardium and an array of biological and/or synthetic materials, there is great potential for any combination of these to assuage the extent of acute ischemic injury and impede the onset of heart failure pursuant to myocardial remodeling. 
Here we present the methods and materials used to reliably perform this microsurgery and the modifications involved for temporary (with reperfusion) or permanent coronary artery occlusion studies as well as intramyocardial injections. The effects on the heart that can be seen during the procedure and at the termination of the experiment in addition to histological evaluation will verify efficacy. 
Briefly, surgical preparation involves anesthetizing the mice, removing the fur on the chest, and then disinfecting the surgical area. Intratracheal intubation is achieved by transesophageal illumination using a fiber optic light. The tubing is then connected to a ventilator. An incision made on the chest exposes the pectoral muscles which will be cut to view the ribs. For ischemia/reperfusion studies, a 1 cm piece of PE tubing placed over the heart is used to tie the ligature to so that occlusion/reperfusion can be customized. For intramyocardial injections, a Hamilton syringe with sterile 30gauge beveled needle is used. When the myocardial manipulations are complete, the rib cage, the pectoral muscles, and the skin are closed sequentially. Line block analgesia is effected by 0.25% marcaine in sterile saline which is applied to muscle layer prior to closure of the skin. The mice are given a subcutaneous injection of saline and placed in a warming chamber until they are sternally recumbent. They are then returned to the vivarium and housed under standard conditions until the time of tissue collection. At the time of sacrifice, the mice are anesthetized, the heart is arrested in diastole with KCl or BDM, rinsed with saline, and immersed in fixative. Subsequently, routine procedures for processing, embedding, sectioning, and histological staining are performed. 
Nonsurgical intubation of a mouse and the microsurgical manipulations described make this a technically challenging model to learn and achieve reproducibility. These procedures, combined with the difficulty in performing consistent manipulations of the ligature for timed occlusion(s) and reperfusion or intramyocardial injections, can also affect the survival rate so optimization and consistency are critical.

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

Enfarktüsü bir murin Model Koroner Arter ligasyon ve intramiyokardiyal Enjeksiyon

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo.  With the advent of genetic modifications to ...

Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related hospitalizations and for which no effective treatments are available to date1,2. Because of the heterogeneity of ICH in humans, various preclinical models are needed to thoroughly explore prospective therapeutic strategies3. Experimental ICH is commonly induced in rodents by intraparenchymal injection of either autologous blood or bacterial collagenase4. The appropriate model is selected based on the pathophysiology of hemorrhage induction and injury progression. The blood injection model mimics a rapidly progressing hemorrhage. Alternatively, bacterial collagenase enzymatically disrupts the basal lamina of brain capillaries, causing an active bleed that generally evolves over several hours5. Resultant perihematomal edema and neurofunctional deficits can be quantified from both models. In this study, we described and evaluated a modified double injection model of autologous whole blood6 as well as an ICH injection model of bacterial collagenase7, both of which target the basal ganglia (corpus striatum) of male CD-1 mice. We assessed neurofunctional deficits and brain edema at 24 and 72 hr after ICH induction. Intrastriatal injection of autologous blood (30 &mu;l) or bacterial collagenase (0.075U) caused reproducible neurofunctional deficits in mice and significantly increased brain edema at 24 and 72 hr after surgery (p&lt;0.05). In conclusion, both models yield consistent hemorrhagic infarcts and represent basic methods for preclinical ICH research.

Clinically relevant animal models of intracerebral hemorrhage (ICH) are needed to extend our knowledge of hemorrhagic stroke and to examine novel therapeutic strategies. In this study, we describe and evaluate two ICH models that implement unilateral injections of either autologous whole blood or bacterial collagenase into the basal ganglia (corpus striatum) of mice.

Farelerde Modelleme İntraserebral Kanama: Otolog Kan veya bakteriyel kollajenaz Enjeksiyon

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

Subaraknoid Kanama bir fare modeli

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

Stereological and Flow Cytometry Characterization of Leukocyte Subpopulations in Models of Transient or Permanent Cerebral Ischemia

Microglia activation, as well as extravasation of haematogenous macrophages and neutrophils, is believed to play a pivotal role in brain injury after stroke. These myeloid cell subpopulations can display different phenotypes and functions and need to be distinguished and characterized to study their regulation and contribution to tissue damage. This protocol provides two different methodologies for brain immune cell characterization: a precise stereological approach and a flow cytometric analysis. The stereological approach is based on the optical fractionator method, which calculates the total number of cells in an area of interest (infarcted brain) estimated by a systematic random sampling. The second characterization approach provides a simple way to isolate brain leukocyte suspensions and to characterize them by flow cytometry, allowing for the characterization of microglia, infiltrated monocytes and neutrophils of the ischemic tissue. In addition, it also details a cerebral ischemia model in mice that exclusively affects brain cortex, generating highly reproducible infarcts with a low rate of mortality, and the procedure for histological brain processing to characterize infarct volume by the Cavalieri method.

Brain myeloid cells characterization following stroke can be performed by stereology using the optical fractionator method, or by a flow cytometric analysis of brain leukocytes suspensions. Both are useful techniques to perform an accurate phenotypical distinction of the main myeloid cell subsets found in the ischemic brain.

Geçici veya kalıcı Serebral İskemi Modelinde lökosit alt popülasyonlarının Stereolojik ve Sitometrisi Karakterizasyonu

Microglia activation, as well as extravasation of haematogenous macrophages and neutrophils, is believed to play a pivotal role in brain injury after ...

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

Subaraknoid Kanama Akut ve Geç sekeli Çalışmaları Tavşan Kan şant Modeli: Teknik Yönleri

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

Ferric Chloride-induced Murine Thrombosis Models

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, diabetes, obesity, cancer and chronic autoimmune rheumatologic disorders. Thrombi are the cause of most heart attacks, strokes and extremity loss, making thrombosis an extremely important public health problem. Since these thrombi stem from inappropriate platelet activation and subsequent coagulation, targeting these systems therapeutically has important clinical significance for developing safer treatments. Due to the complexities of the hemostatic system, in vitro experiments cannot replicate the blood-to-vessel wall interactions; therefore, in vivo studies are critical to understand pathological mechanisms of thrombus formation. To this end, various thrombosis models have been developed in mice. Among them, ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation and aggregation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for the development of a thrombus that occludes blood flow gives a quantitative measure of vascular injury, platelet activation and aggregation that is relevant to thrombotic diseases. We have significantly refined this FeCl3-induced vascular thrombosis model, which makes the data highly reproducible with minimal variation. Here we describe the model and present representative data from several experimental set-ups that demonstrate the utility of this model in thrombosis research.

We report a refined procedure of the ferric chloride (FeCl3)-induced thrombosis models on carotid and mesenteric artery as well as vein, characterized efficiently using intravital microscopy to monitor time to occlusive thrombi formation.

Ferrik klorit indüklenen fare tromboz modellerinde

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, ...

Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflammatory Bowel Diseases

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have immunosuppressive potential and have been shown to suppress inflammation in mouse models of intestinal inflammation, even though the route of administration can limit their homing and effectiveness 1,3,4,5. Local application of MSCs to colonic injury models has shown greater efficacy at ameliorating inflammation in the colon. However, there is paucity of data on techniques to enhance the localization of human bone marrow-derived MSCs (hMSCs) to the small intestine, the site of inflammation in the SAMP-1/YitFc (SAMP) model of experimental Crohn&#39;s disease. This work describes a novel technique for the ultrasound-guided intracardiac injection of hMSCs in SAMP mice, a well-characterized spontaneous model of chronic intestinal inflammation. Sex- and age-matched, inflammation-free AKR/J (AKR) mice were used as controls. To analyze the biodistribution and the localization, hMSCs were transduced with a lentivirus containing a triple reporter. The triple reporter consisted of firefly luciferase (fl), for bioluminescent imaging; monomeric red fluorescent protein (mrfp), for cell sorting; and truncated herpes simplex virus thymidine kinase (ttk), for positron emission tomography (PET) imaging. The results of this study show that 24 h after the intracardiac administration, hMSCs localize in the small intestine of SAMP mice as opposed to inflammation-free AKR mice. This novel, ultrasound-guided injection of hMSCs in the left ventricle of SAMP mice ensures a high success rate of cell delivery, allowing for the rapid recovery of mice with minimal morbidity and mortality. This technique could be a useful method for the enhanced localization of MSCs in other models of small-intestinal inflammation, such as TNF&#916;RE6. Future studies will determine if the increased localization of hMSCs by intra-arterial delivery can lead to increased therapeutic efficacy.

Murine studies in models of colonic inflammation have demonstrated that a small percentage (1 - 5%) of mesenchymal stem cells (MSC) injected intravenously or intraperitoneally home to the inflamed colon1,2. This study shows that ultrasound-guided intracardiac injections of MSCs result in increased localization to the intestine.

Ultrason destekli İntrakardiyak enjeksiyon bağırsak için giriş için deneysel iltihabi bağırsak hastalıkları fare modellerinde kullanım artırmak için insan Mezenkimal Kök hücre

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have ...

Nerve Stimulator-guided Injection of Autologous Stem Cells Near the Equine Left Recurrent Laryngeal Nerve

Recurrent laryngeal neuropathy (RLN) commonly affects horses and is characterized by abnormal respiratory sounds and exercise intolerance. The recurrent laryngeal nerve shows lesions of demyelination. The benefit of applying stem cells to demyelinated nerves has been demonstrated in various animal models. The aim of the study was to test the feasibility and safety of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to the left recurrent laryngeal nerve in healthy horses by using an electrical nerve stimulator.
Muscle-derived stems cell are obtained from five healthy Standardbred horses by sampling 20 mg of muscle tissue with a semi-automatic 14 G biopsy needle from the triceps muscle. Movements of the larynx are monitored via upper-airway video endoscopy. The left recurrent laryngeal nerve is approached with an insulated nerve block needle. Nerve stimulation is applied, starting at 2 mA, and the successful abduction of the left arytenoid is monitored. The stimulation intensity is reduced progressively. When a loss of the motor response is observed at 0.5 mA, 107 autologous muscle-derived stem cells are injected. Two examiners, who are blinded to the time point, score the laryngeal function of the horses prior to the treatment and at day 1, day 7, and day 28 after the injection of the cells. In a sixth horse, 1 mL of 2% lidocaine is injected to further confirm the correct positioning of the needle. This leads to a temporary paralysis of the left arytenoid cartilage.
This study proves that the recurrent laryngeal nerve can be approached with the help of an electrical nerve stimulator and that the electrical stimulation of the nerve is well tolerated by the horses. No modification of the laryngeal function was observed in any of the horses after the injection of the stem cells. Further studies should be conducted to describe the effects of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to horses suffering from RLN.

Here, we present a protocol to inject autologous muscle-derived stem cells in the proximity to the recurrent laryngeal nerve with the help of an electrical nerve stimulator. This novel technique may become useful for the treatment of equine recurrent laryngeal neuropathy.

Otolog kök hücre yakınındaki at sol tekrarlayan gırtlak sinir sinir Stimülatörü güdümlü enjeksiyon

Recurrent laryngeal neuropathy (RLN) commonly affects horses and is characterized by abnormal respiratory sounds and exercise intolerance. The recurrent ...

Subclavian Vein Puncture As an Alternative Method of Blood Sample Collection in Rats

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful compared to other more aggressive methods such as retro-orbital plexus sample collection. However, this blood collection method is sometimes limited by an unsatisfactory success rate. Here, we introduce a method for blood collection through the subclavian vein puncture. The subclavian vein is located just under the clavicle and this vein is large enough to fulfill the volume requirements of blood collection. Our results show that this method is safe and applicable for blood collection sampling with the required blood volume. Blood collection through the subclavian vein puncture could serve as an alternative blood collection method in case of failed tail vein blood sampling in rats.

Here, we present a protocol to collect blood samples from the rat subclavian vein.

Subklaviyan ven ponksiyon kan örnek fareleri koleksiyonunda alternatif bir yöntem olarak

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful ...