Bu araştırma DZHK (Almanca merkezi kardiyovasküler araştırma, DB), tarafından desteklenmiştir (Else-Kröner-Fresenius Stiftung, F.H.), EKFS ve BMBF (Alman Milli Eğitim Bakanlığı ve araştırma), hem de klinik bilim adamı programı finanse BIH Charité tarafından Charité tarafından - Universitätsmedizin Berlin ve Berlin Sağlığı Enstitüsü (F.H.).

<ol>
	<li>Alberti, K. 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=Harmonizing+the+metabolic+syndrome:+a+joint+interim+statement+of+the+International+Diabetes+Federation+Task+Force+on+Epidemiology+and+Prevention;+National+Heart,+Lung,+and+Blood+Institute;+American+Heart+Association;+World+Heart+Federation;+International+Atherosclerosis+Society;+and+International+Association+for+the+Study+of+Obesity.">Harmonizing the metabolic syndrome: a joint interim statement of the International Diabetes Federation Task Force on Epidemiology and Prevention; National Heart, Lung, and Blood Institute; American Heart Association; World Heart Federation; International Atherosclerosis Society; and International Association for the Study of Obesity.</a> Circulation. 120 (16), 1640-1645 (2009).</li><li>. IDF Consensus Worldwide Definition of the Metabolic Syndrome Available from: <a target="_blank" href="https://www.idf.org/e-library/consensus-statements/60-idfconsensus-worldwide-definitionof-the-metabolic-syndrome.html">https://www.idf.org/e-library/consensus-statements/60-idfconsensus-worldwide-definitionof-the-metabolic-syndrome.html</a> (2006)</li><li>Melenovsky, V., 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=Left+atrial+remodeling+and+function+in+advanced+heart+failure+with+preserved+or+reduced+ejection+fraction.">Left atrial remodeling and function in advanced heart failure with preserved or reduced ejection fraction.</a> Circulation: Heart Failure. 8 (2), 295-303 (2015).</li><li>Goette, 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=EHRA/HRS/APHRS/SOLAECE+expert+consensus+on+atrial+cardiomyopathies:+definition,+characterization,+and+clinical+implication.">EHRA/HRS/APHRS/SOLAECE expert consensus on atrial cardiomyopathies: definition, characterization, and clinical implication.</a> EP Europace. 18 (10), 1455-1490 (2016).</li><li>Schotten, U., Verheule, S., Kirchhof, P., Goette, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiological+mechanisms+of+atrial+fibrillation:+a+translational+appraisal.">Pathophysiological mechanisms of atrial fibrillation: a translational appraisal.</a> Physiological Reviews. 91 (1), 265-325 (2011).</li><li>Hohendanner, F., DeSantiago, J., Heinzel, F. 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Yazarlar ifşa gerek yok.

Burada, ilk tek hücreli ACMs atriyal remodeling22işaretlenmiş gösteren MetS ile ilgili HFpEF bir fare modeli yalıtım için bir protokol açıklanan. Aşırı yağ dokusu aort, giderek daha zor cannulation yanı sıra cerrahi hazırlık yapabilir gibi benzersiz olarak zor bir işlemdir. Sorun giderme kılavuzu Tablo 2 adresleri yalıtım yordam en yaygın sorunların sağlanan.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbod...

MetS tip-2 diyabet ve kalp-damar hastalıkları için risk faktörlerinin bir küme tanımlar ve artan bir arteryel kan basıncı içerir (trigliserid yükseltilmiş ve alçaltılmış yüksek yoğunluklu lipoprotein kolesterol) dislipidemi, artan açlık glikoz ve merkezi obezite1. MetS dünya çapında yaygınlık % 25-30 ve sürekli yükselen2olduğu tahmin edilmektedir. HFpEF hànzú bir klinik sendrom genellikle MetS ile ilgili olduğunu. HFpEF ve onun önceki aşamaları (yani, hipertansif kalp hastalığı) sırasında kalp remodeling da kulakçık3/ remodeling tarafından eşlik eder. İndirimli contractile işlevi ve sol atriyum yapısal değişiklikler artmış mortalite, atriyal fibrilasyon ve yeni başlangıçlı kalp yetmezliği4ile ilişkili bulunmuştur. Atriyal remodeling iyon kanal işlevi, Ca2 + homeostazı, atriyal yapısı, fibroblast harekete geçirmek ve doku fibrozu5değişiklikler karakterizedir. Atriyal MetS ile ilgili HFpEF ve onun altında yatan patolojik mekanizmaları hala kötü anlaşılır ve bir daha da derinlemesine araştırma gereken remodeling için yorum yaptı. Hayvan modelleri var olmak a değerli alet ve atriyal kardiyomiyopatiler6,7,8,9alanında birçok gelişmeler yol kanıtlanmıştır.
Çalışmalar izole tek hücreli cardiomyocytes ile teyit ve in vivo bulgular tamamlayıcı için sık sık kullanılır. Bir izolasyon ve potansiyel sonraki hücre kültürü, yollar, iyonik kanal akımları ve uyarma-kasılma-kaplin sinyal soruşturma için izin verir. Fizyolojik koşullar altında cardiomyocytes çoğalırlar değil. Füzyon bir atrial natriuretic faktör transkripsiyon düzenleyici dizisi ve bir simian virüs 40 büyük T antijen transgenik farelerde arasında AT-110adlı ilk ölümsüzleştirdi ACMs yol açtı. AT-1 hücreleri daha da geliştirilmesi sadece seri olarak passaged edemem ama aynı zamanda kendiliğinden sözleşme11HL-1 hücrelere doğuran. Ancak, daha az organize ultrastructure myofibrils11ve bir hyperpolarization harekete geçirmek içe geçerli12geliştirme yüksek bir olay gibi taze izole hücrelere kıyasla yapısal ve işlevsel farklılıklar göstermektedir. Ventrikül cardiomyocytes (VCM) sıçanlar ve fareler modelleri çeşitli yalıtım iyi kurulmuş13,14,15,16,17,18 ' dir , 19. genel olarak eksize kalp Langendorff aparatı monte ve retrogradely bir Ca2 +ile derin-collagenases ve proteaz gibi sindirim enzimleri içeren boş arabellek. Kalsiyum o zaman fizyolojik şartlarda kademeli bir şekilde yeniden ortaya çıkar. Ancak, kullanışlılığı hastalık modelleri atriyal remodeling ile sınırlı olmasına rağmen ACMs yalıtım için adanmış protokolleri kullanılabilir20,21, artan fibrozis ve basınç ilgili farklar nedeniyle.
Bu makalede, bir protokol atriyal remodeling (yani, özellikle ZFS1 fare modeli MetS ile ilgili HFpEF için)22göstermek hayvanlardan atriyal tek hücreli cardiomyocytes yalıtım için hayata geçirdik. Varolan yalıtım protokolleri en iyi duruma getirilmiş ve kontrol ve morfolojik ve fonksiyonel olarak sağlam cardiomyocytes daha yüksek verimleri için önde gelen kardiyak boşluklar, intraluminal basınç değiştirmek için basit, özel yapım bir aygıt tarafından tamamlanmaktadır. Aşağıdaki iletişim kuralı bir adım adım kılavuz ile araştırmacı, özel yapım donanımları, bir liste-in eriyik yanı sıra kapsamlı sorun giderme kılavuzu ayrıntılı bir açıklamasını sağlar.

<table border="0" cellpadding="0" cellspacing="0" width="935">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">ZSF-1 Obese rat</td>
			<td style="width:292px;">Charles River Laboratories, Inc.</td>
			<td style="width:136px;"></td>
			<td style="width:227px;">21 weeks old</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Fine Iris Scissors</td>
			<td style="width:292px;">Fine Science Tools GmbH</td>
			<td style="width:136px;">14094-11</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Surgical Scissors</td>
			<td style="width:292px;">Fine Science Tools GmbH</td>
			<td style="width:136px;">14001-18</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Micro Dressing Forceps (curved, serrated)</td>
			<td style="width:292px;">Aesculap, Inc.</td>
			<td style="width:136px;">BD312R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Tissue Forceps (straight, 1 x 2 teeth)</td>
			<td style="width:292px;">Aesculap, Inc.</td>
			<td style="width:136px;">BD537R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Tying Forceps (angled)</td>
			<td style="width:292px;">Aesculap, Inc.</td>
			<td style="width:136px;">MA624R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Rodent and Small Animal Guillotine</td>
			<td style="width:292px;">Kent Scientific Corp.</td>
			<td style="width:136px;">DCAP</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Low Cost Induction Chamber 3.0 L</td>
			<td style="width:292px;">Kent Scientific Corp.</td>
			<td style="width:136px;">SOMNO-0730&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Butterfly Winged Infusion Set 21 G</td>
			<td style="width:292px;">Hospira, Inc.</td>
			<td style="width:136px;">181106101</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Abbocath 16 G</td>
			<td style="width:292px;">Hospira, Inc.</td>
			<td>0G7149702</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Microlance Hypodermic Needle</td>
			<td style="width:292px;">Becton Dickinson GmbH</td>
			<td style="width:136px;">301300</td>
			<td>modify needle to make cannula</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Braun Original Perfusor Syringe 50 ml</td>
			<td style="width:292px;">B. Braun Melsungen AG</td>
			<td style="width:136px;">8728810F</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Braun Inject Solo Syringe 10 ml</td>
			<td style="width:292px;">B. Braun Melsungen AG</td>
			<td style="width:136px;">2057926</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Beaker 50ml</td>
			<td style="width:292px;">Duran Group (DWK Life Sciences GmbH)</td>
			<td>21 106 17</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Duroplan petri dish (100 x 20 mm)</td>
			<td style="width:292px;">Duran Group (DWK Life Sciences GmbH)</td>
			<td style="width:136px;">21 755 48</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Seraflex Suture USP 3/0</td>
			<td style="width:292px;">SERAG-WIESSNER GmbH &#38; Co. KG</td>
			<td style="width:136px;">IC208000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">VWR disposable Square Weighin Boats 100ml</td>
			<td style="width:292px;">VWR, Inc.</td>
			<td style="width:136px;">10803-148</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Styrofoam surface</td>
			<td style="width:292px;"></td>
			<td style="width:136px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Sodium chloride</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">71380</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Potassium chloride</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">P4504</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Potassium phosphate monobasic</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">P5379</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Sodium phosphate dibasic</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">S0876</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Magensium sulfate heptahydrate</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">230391</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Magensium chloride</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">M8266</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">HEPES</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">H3375</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Taurine</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">T0625</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Glucose</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">G7528</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">2,3-Butanedione monoxime</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">B0753</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Calcium chloride solution (1 M)</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">21115</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Bovine Serum Albumin</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">A9647</td>
			<td style="width:227px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Liberase</td>
			<td style="width:292px;">Roche (Sigma-Aldrich, Inc.)</td>
			<td style="width:136px;">LIBTM-RO</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Heparin</td>
			<td style="width:292px;">Rotexmedica GmbH</td>
			<td style="width:136px;">3862357</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Forene (Isoflurane)</td>
			<td style="width:292px;">Abbvie Deutschland GmbH &#38; Co. KG</td>
			<td style="width:136px;">10182054</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane</td>
			<td style="width:292px;">Sigma-Aldrich, Inc.</td>
			<td style="width:136px;">L2020</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">WillCo glass-bottom dish 500&#181;l 0.005mm</td>
			<td style="width:292px;">WillCo Wells B.V.</td>
			<td style="width:136px;">HBST-3522</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Fluo4 AM</td>
			<td style="width:292px;">Invitrogen (Thermo Fisher Scientific, Inc.)</td>
			<td>F14201</td>
			<td>5&#181;M for 20min at RT</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Di-8-ANNEPS</td>
			<td style="width:292px;">Invitrogen (Thermo Fisher Scientific, Inc.)</td>
			<td>D3167</td>
			<td>10&#181;M for 45 min at 37&#176; C&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Mitotracker RED FM</td>
			<td style="width:292px;">Invitrogen (Thermo Fisher Scientific, Inc.)</td>
			<td style="width:136px;">M22425</td>
			<td>20nM for 30 min at 37&#176; C</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Jacketed reaction vessel 500 ml</td>
			<td style="width:292px;">Gebr. Rettberg GmbH</td>
			<td>107024414</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Jacketed reaction vessel 1000 ml</td>
			<td style="width:292px;">Gebr. Rettberg GmbH</td>
			<td>107025414</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Jacketed bubble trap</td>
			<td style="width:292px;">Gebr. Rettberg GmbH</td>
			<td>134720001</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">ED heating immersion circulator</td>
			<td style="width:292px;">Julabo GmbH</td>
			<td>9116000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Reglo Digital MS-2/6 peristaltic pump</td>
			<td style="width:292px;">Ismatec (Cole-Parmer Gmbh)</td>
			<td style="width:136px;">ISM 831</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Voltcraft Thermometer 302 K/J</td>
			<td style="width:292px;">Conrad Electronic SE</td>
			<td style="width:136px;">030300546</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Tubing</td>
			<td style="width:292px;"></td>
			<td style="width:136px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">LSM 700 microscope</td>
			<td style="width:292px;">Carl Zeiss, Inc.</td>
			<td style="width:136px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">ZEN 2.3 imaging software</td>
			<td style="width:292px;">Carl Zeiss, Inc.</td>
			<td style="width:136px;">410135-1011-240&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">Single channel heater controller TC-324B</td>
			<td style="width:292px;">Warner Instruments, LLC</td>
			<td style="width:136px;">64-2400</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">8 channel perfusion system</td>
			<td style="width:292px;">Warner Instruments, LLC</td>
			<td style="width:136px;">64-0185</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:281px;">8 channel Multi-Line In-Line Solution Heaters</td>
			<td style="width:292px;">Warner Instruments, LLC</td>
			<td style="width:136px;">64-0105</td>
			<td></td>
		</tr>
	</tbody>
</table>

isolation of atrial cardiomyocytes from a rat model of metabolic syndrome-related heart failure with preserved ejection fraction

Bu makalede, biz bir en iyi duruma getirilmiş, Langendorff tabanlı tek hücreli atriyal cardiomyocytes (ACMs) bir fare modeli Metabolik sendromu (MetS) yalıtım için açıklayınız-kalp yetmezliği korunmuş ejeksiyon fraksiyonu (HFpEF) ile ilgili. MetS ile ilgili HFpEF yaygınlığı artıyor ve atriyal remodeling mortalite bağımsız bir tahmin olarak atriyal kardiyomiyopatiler atriyal remodeling ve atriyal fibrilasyon ile ilgili klinik olarak son derece ilgilidir. Çalışmalar izole tek hücreli cardiomyocytes ile teyit ve in vivo bulgular tamamlayıcı için sık sık kullanılır. Dolaşım gemi rarefication ve interstisyel doku fibrozu ACMs başarılı tek hücreli yalıtım bu hastalığın hayvan modeller için potansiyel olarak kısıtlayıcı bir faktör poz.
Biz bu sorunu bir aygıtı el ile yalıtım işlem sırasında kalp boşlukları intraluminal basınç düzenleyen yetenekli istihdam ederek önemli ölçüde morfolojik ve fonksiyonel olarak sağlam ACMs verimini artırma ele sahip. Alınan hücreler, hücre kültürü ve fonksiyonel kalsiyum görüntüleme (yani, uyarma-kasılma-kaplin) gibi farklı deneyler çeşitli kullanılabilir.
Araştırmacı bir adım adım protokolüyle, listesini en iyi duruma getirilmiş çözümler, gerekli ekipman ve kapsamlı sorun giderme Rehberi hazırlamak için ayrıntılı yönergeler sağlar. Yordamın ilk uygulama oldukça zor olabilir, başarılı bir uyum okuyucu MetS ile ilgili HFpEF deneyler geniş bir yelpazede, fare modeli state-of--art ACM izolasyonların gerçekleştirmek izin verir.

21 haftalıkken 60-90 oranında kalsiyum yeniden uyarlaması (adım 5.4-5,7), (adım 6.1 açıklandığı gibi tahmini) uygun ACMs ZSF-1 obez sıçanlarından emir bu Yöntem (Şekil 4A) tarafından ayrılmış olabilir. Rat, farklı bir ve daha fazla hànzú fenotip VCMs24,25ile karşılaştırıldığında ACMs karakterizedir. Şekil 4B korunmuş membranlar ve sarcomere...

Watch this Scientific Journal Video about Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction at JoVE.com

Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction

Bir fare modeli Metabolik sendromu ile ilgili kalp yetmezliği ile korunmuş ejeksiyon fraksiyonu atriyal Cardiomyocytes izolasyonu

Burada, biz bir en iyi duruma getirilmiş, Langendorff tabanlı bir fare modeli Metabolik sendromu ile ilgili kalp yetmezliği ile korunmuş ejeksiyon fraksiyonu tek hücreli atriyal cardiomyocytes yalıtım için açıklayınız. El ile düzenleme intraluminal basınç kalp boşlukları, işlevsel olarak sağlam miyositler uyarma-kasılma-kaplin çalışmaları için uygun verim için uygulanır.

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of ...

isolation-atrial-cardiomyocytes-from-rat-model-metabolic-syndrome

Research

JoVE Journal

Medicine

10.0K Görüntüleme.  Charité University Medicine. Burada, biz bir en iyi duruma getirilmiş, Langendorff tabanlı bir fare modeli Metabolik sendromu ile ilgili kalp yetmezliği ile korunmuş ejeksiyon fraksiyonu tek hücreli atriyal cardiomyocytes yalıtım için açıklayınız. El ile düzenleme intraluminal basınç kalp boşlukları, işlevsel olarak sağlam miyositler uyarma-kasılma-kaplin çalışmaları için uygun verim için uygulanır.

Video: Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

Klinik öncesi Model İskemik Kalp Yetmezliği Gen Transferi

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

Tıp

The WATCHMAN Left Atrial Appendage Closure Device for Atrial Fibrillation

Atrial fibrillation (AF) is the most common cardiac arrhythmia, affecting an estimated 6 million people in the United States 1. Since AF affects primarily elderly people, its prevalence increases parallel with age. As such, it is expected that 15.9 million Americans will be affected by the year 2050 2. Ischemic stroke occurs in 5% of non-anticoagulated AF patients each year. Current treatments for AF include rate control, rhythm control and prevention of stroke 3.
The American College of Cardiology, American Heart Association, and European Society of Cardiology currently recommended rate control as the first course of therapy for AF 3. Rate control is achieved by administration of pharmacological agents, such as &beta;-blockers, that lower the heart rate until it reaches a less symptomatic state 3. Rhythm control aims to return the heart to its normal sinus rhythm and is typically achieved through administration of antiarrhythmic drugs such as amiodarone, electrical cardioversion or ablation therapy. Rhythm control methods, however, have not been demonstrated to be superior to rate-control methods 4-6. In fact, certain antiarrhythmic drugs have been shown to be associated with higher hospitalization rates, serious adverse effects 3, or even increases in mortality in patients with structural heart defects 7. Thus, treatment with antiarrhythmics is more often used when rate-control drugs are ineffective or contraindicated. Rate-control and antiarrhythmic agents relieve the symptoms of AF, including palpitations, shortness of breath, and fatigue 8, but don't reliably prevent thromboembolic events 6.
Treatment with the anticoagulant drug warfarin significantly reduces the rate of stroke or embolism 9,10. However, because of problems associated with its use, fewer than 50% of patients are treated with it. The therapeutic dose is affected by drug, dietary, and metabolic interactions, and thus requires detailed monitoring. In addition, warfarin has the potential to cause severe, sometimes lethal, bleeding 2. As an alternative, aspirin is commonly prescribed. While aspirin is typically well tolerated, it is far less effective at preventing stroke 10. Other alternatives to warfarin, such as dabigatran 11 or rivaroxaban 12 demonstrate non-inferiority to warfarin with respect to thromboembolic events (in fact, dabigatran given as a high dose of 150 mg twice a day has shown superiority). While these drugs have the advantage of eliminating dietary concerns and eliminating the need for regular blood monitoring, major bleeding and associated complications, while somewhat less so than with warfarin, remain an issue 13-15.
Since 90% of AF-associated strokes result from emboli that arise from the left atrial appendage (LAA) 2, one alternative approach to warfarin therapy has been to exclude the LAA using an implanted device to trap blood clots before they exit. Here, we demonstrate a procedure for implanting the WATCHMAN Left Atrial Appendage Closure Device. A transseptal cannula is inserted through the femoral vein, and under fluoroscopic guidance, inter-atrial septum is crossed. Once access to the left atrium has been achieved, a guidewire is placed in the upper pulmonary vein and the WATCHMAN Access Sheath and dilator are advanced over the wire into the left atrium. The guidewire is removed, and the access sheath is carefully advanced into the distal portion of the LAA over a pigtail catheter. The WATCHMAN Delivery System is prepped, inserted into the access sheath, and slowly advanced. The WATCHMAN device is then deployed into the LAA. The device release criteria are confirmed via fluoroscopy and transesophageal echocardiography (TEE) and the device is released.

The accompanying video describes a procedure for percutaneous placement of the WATCHMAN Left Atrial Appendage (LAA) Device. The WATCHMAN is a nitinol device designed to be permanently implanted at, or slightly distal to, the opening of the left atrial appendage (LAA) to trap blood clots before they exit the LAA, preventing thromboembolic stroke.

Bekçi Atriyal fibrilasyon için Atriyal Apendiks Kapatma Cihaz Sol

Atrial fibrillation (AF) is the most common cardiac arrhythmia, affecting an estimated 6 million people in the United States 1. Since AF affects primarily ...

A New Single Chamber Implantable Defibrillator with Atrial Sensing: A Practical Demonstration of Sensing and  Ease of Implantation

Implantable cardioverter-defibrillators (ICDs) terminate ventricular tachycardia (VT) and ventricular fibrillation (VF) with high efficacy and can protect patients from sudden cardiac death (SCD). However, inappropriate shocks may occur if tachycardias are misdiagnosed. Inappropriate shocks are harmful and impair patient quality of life. The risk of inappropriate therapy increases with lower detection rates programmed in the ICD. Single-chamber detection poses greater risks for misdiagnosis when compared with dual-chamber devices that have the benefit of additional atrial information. However, using a dual-chamber device merely for the sake of detection is generally not accepted, since the risks associated with the second electrode may outweigh the benefits of detection. Therefore, BIOTRONIK developed a ventricular lead called the LinoxSMART S DX, which allows for the detection of atrial signals from two electrodes positioned at the atrial part of the ventricular electrode. This device contains two ring electrodes; one that contacts the atrial wall at the junction of the superior vena cava (SVC) and one positioned at the free floating part of the electrode in the atrium. The excellent signal quality can only be achieved by a special filter setting in the ICD (Lumax 540 and 740 VR-T DX, BIOTRONIK). Here, the ease of implantation of the system will be demonstrated.

Dual-chamber implantable cardioverter-defibrillators (ICDs) may improve detection of atrial fibrillation as well as differentiation of tachycardias. However, this advantage is undermined by complications associated with the second electrode, which is required in conventional dual chamber devices. Therefore, BIOTRONIK has developed a new electrode called the LinoxSMART S DX that, when used in conjunction with the Lumax DX ICD, offers dual-chamber detection without the risks associated with the second electrode.

Yeni Bir Tek Odası İmplante Atriyal Algılama ile Defibrilatör: Algılama Pratik Bir Gösteri ve İmplantasyon kolaylığı

Implantable cardioverter-defibrillators (ICDs) terminate ventricular tachycardia (VT) and ventricular fibrillation (VF) with high efficacy and can protect ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

İzole Yöntemi<em&gt; Ex vivo</emBir Rat Modelinde&gt; Akciğer Perfüzyon: Sıçan EVLP Programı Geliştirme Öğrenilmiş Dersler

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart

The present protocol describes a unique approach that enables the collection of cardiac transudate (CT) from the isolated, saline-perfused rat heart. After isolation and retrograde perfusion of the heart according to the Langendorff technique, the heart is inverted into an upside-down position and is mechanically stabilized by a balloon catheter inserted into the left ventricle. Then, a thin latex cap &#8211; previously cast to match the average size of the rat heart &#8211; is placed over the epicardial surface. The outlet of the latex cap is connected to silicon tubing, with the distal opening 10 cm below the base level of the heart, creating slight suction. CT continuously produced on the epicardial surface is collected in ice-cooled vials for further analysis. The rate of CT formation ranged from 17 to 147 &#181;L/min (n = 14) in control and infarcted hearts, which represents 0.1-1% of the coronary venous effluent perfusate. Proteomic analysis and high performance liquid chromatography (HPLC) revealed that the collected CT contains a wide spectrum of proteins and purinergic metabolites.

This protocol describes a method to collect cardiac interstitial fluid from the isolated, perfused rat heart. To physically separate interstitial transudate from coronary venous effluent perfusate, the Langendorff perfused heart is inverted, and the transudate (interstitial fluid) formed on the cardiac surface is collected using a soft latex cap.

İzole edilen fare kalpten interstisyel transüdao toplanması için ters kalp modeli

The present protocol describes a unique approach that enables the collection of cardiac transudate (CT) from the isolated, saline-perfused rat heart. ...

Isolation and Cultivation of Adult Rat Cardiomyocytes

In an intact heart, adjacent cells influence adult cardiomyocytes. With the method of isolation and cultivation of adult cardiomyocytes, a precise investigation of the behavior of these cells under specific treatments and environments is possible. This manuscript presents a protocol for successful isolation and cultivation of adult rat ventricular cardiomyocytes (ARVC).
The rat is sacrificed by cervical dislocation under deep anesthesia. Then, the heart is extracted and the aorta is uncovered. Subsequently, perfusion on the Langendorff perfusion system with calcium depletion and collagenase treatment is performed. Afterwards, ventricular tissue gets minced, re-circulated, and filtered, followed by three centrifugation steps with gradual addition of CaCl2 until physiological calcium concentration is reached. ARVC are plated on cell culture dishes. After refreshing the cell culture medium, ARVC can be cultivated for up to six days without changing the serum-containing culture medium. Isolation of ARVC is a calcium sensitive process. Small changes in the intracellular calcium concentration cause a decrease in the quality and viability of the isolated cells.
Freshly isolated ARVC are rod shaped. Within the first days of cultivation they lose the rod-shaped morphology and form pseudopodia-like structures (spreading). During this morphological formation ARVC initially degrade their contractile elements followed by a reformation through actin stress fibers and de novo sarcomerogenesis. After one week of cultivation, most ARVC show a widespread appearance with a clearly detectable cross striation. This process is sensitive to intracellular calcium concentration, as treatment with ionomycin attenuates spreading. Key markers in this process of de- and re-differentiation are &#946;-myosin heavy chain (&#946;-MHC), oncostatin M (OSM), and swiprosin-1 (EFHD2). Recent studies have suggested that cardiac re- and de-differentiation occurring under culture conditions mimics features seen in vivo during cardiac remodeling. Therefore, isolation and cultivation of ARVC play a key role in understanding the biology of cardiomyocytes.

Here, we present a protocol for the isolation and cultivation of adult rat ventricular cardiomyocytes (ARVC). Isolated ARVC can be used for short and long-term cultivation. The isolation and cultivation of ARVC can play a key role in developing new treatment regimens for cardiac diseases.

Yalıtım ve yetişkin fare Cardiomyocytes tarımı

In an intact heart, adjacent cells influence adult cardiomyocytes. With the method of isolation and cultivation of adult cardiomyocytes, a precise ...

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment methods. Here, we present such a model by tachycardia-induced cardiomyopathy, which can be produced by rapid cardiac pacing in swine.
A single pacing lead is introduced transvenously into fully anaesthetized healthy swine, to the apex of the right ventricle, and fixated. Its other end is then tunneled dorsally to the paravertebral region. There, it is connected to an in-house modified heart pacemaker unit that is then implanted in a subcutaneous pocket. 
After 4 - 8 weeks of rapid ventricular pacing at rates of 200 - 240 beats/min, physical examination revealed signs of severe heart failure - tachypnea, spontaneous sinus tachycardia, and fatigue. Echocardiography and X-ray showed dilation of all heart chambers, effusions, and severe systolic dysfunction. These findings correspond well to decompensated dilated cardiomyopathy and are also preserved after the cessation of pacing.
This model of tachycardia-induced cardiomyopathy can be used for studying the pathophysiology of progressive chronic heart failure, especially hemodynamic changes caused by new treatment modalities like mechanical circulatory supports. This methodology is easy to perform and the results are robust and reproducible.

Here, we present a protocol to produce tachycardia-induced cardiomyopathy in swine. This model represents a potent way to study the hemodynamics of progressive chronic heart failure and the effects of applied treatment.

Kronik kalp yetmezliği model domuz olarak taşikardi kaynaklı kardiyomiyopati

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

Isolation of Atrial Myocytes from Adult Mice

The electrophysiological properties of atrial myocytes importantly affect overall cardiac function. Alterations in the underlying ionic currents responsible for the action potential can cause pro-arrhythmic substrates that underlie arrhythmias, such as atrial fibrillation, which are highly prevalent in many conditions and disease states. Isolating adult mouse atrial cardiomyocytes for use in patch-clamp experiments has greatly advanced our knowledge and understanding of the cellular electrophysiology in the healthy atrial myocardium and in the setting of atrial pathophysiology. In addition, studies using genetic mouse models have elucidated the role of a vast array of proteins in regulating atrial electrophysiology. Here we provide a detailed protocol for the isolation of cardiomyocytes from the atrial appendages of adult mice using a combination of enzymatic digestion and mechanical dissociation of these tissues. This approach consistently and reliably yields isolated atrial cardiomyocytes that can then be used to characterize cellular electrophysiology by measuring action potentials and ionic currents in patch-clamp experiments under a number of experimental conditions.

This protocol is used to isolate single atrial cardiomyocytes from the adult mouse heart using a chunk digestion approach. This approach is used to isolate right or left atrial myocytes that can be used to characterize atrial myocyte electrophysiology in patch-clamp studies.

Yetişkin fareler atriyal Miyoksit izolasyonu

The electrophysiological properties of atrial myocytes importantly affect overall cardiac function. Alterations in the underlying ionic currents ...

Establishing a Swine Model of Post-myocardial Infarction Heart Failure for Stem Cell Treatment

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major causes of mortality and morbidity around the world. Cell-based therapies for cardiac repair and improvement of left ventricular function after MI have attracted considerable attention. Accordingly, the safety and efficacy of these cell transplantations should be tested in a preclinical large animal model of HF prior to clinical use. Pigs are widely used for cardiovascular disease research due to their similarity to humans in terms of heart size and coronary anatomy. Therefore, we sought to present an effective protocol for the establishment of a porcine chronic HF model using closed-chest coronary balloon occlusion of the left circumflex artery (LCX), followed by rapid ventricular pacing induced with pacemaker implantation. Eight weeks later, the stem cells were administered by intramyocardial injection in the peri-infarct area. Then the infarct size, cell survival, and left ventricular function (including echocardiography, hemodynamic parameters, and electrophysiology) were evaluated. This study helps establish a stable preclinical large animal HF model for stem cell treatment.

We sought to establish a swine model of heart failure induced by left circumflex artery blockage and rapid pacing to test the effect and safety of intramyocardial administration of stem cells for cell-based therapies.

Kök Hücre Tedavisi Için Post-miyokard Enfarktüsü Kalp Yetmezliği Domuz Modeli Oluşturma

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major ...

A Rat Model of Pressure Overload Induced Moderate Remodeling and Systolic Dysfunction as Opposed to Overt Systolic Heart Failure

In response to an injury, such as myocardial infarction, prolonged hypertension or a cardiotoxic agent, the heart initially adapts through the activation of signal transduction pathways, to counteract, in the short-term, for the cardiac myocyte loss and or the increase in wall stress. However, prolonged activation of these pathways becomes detrimental leading to the initiation and propagation of cardiac remodeling leading to changes in left ventricular geometry and increases in left ventricular volumes; a phenotype seen in patients with systolic heart failure (HF). Here, we describe the creation of a rat model of pressure overload induced moderate remodeling and early systolic dysfunction (MOD) by ascending aortic banding (AAB) via a vascular clip with an internal area of 2 mm2. The surgery is performed in 200 g Sprague-Dawley rats. The MOD HF phenotype develops at 8-12 weeks after AAB and is characterized noninvasively by means of echocardiography. Previous work suggests the activation of signal transduction pathways and altered gene expression and post-translational modification of proteins in the MOD HF phenotype that mimic those seen in human systolic HF; therefore, making the MOD HF phenotype a suitable model for translational research to identify and test potential therapeutic anti-remodeling targets in HF. The advantages of the MOD HF phenotype compared to the overt systolic HF phenotype is that it allows for the identification of molecular targets involved in the early remodeling process and the early application of therapeutic interventions. The limitation of the MOD HF phenotype is that it may not mimic the spectrum of diseases leading to systolic HF in human. Moreover, it is a challenging phenotype to create, as the AAB surgery is associated with high mortality and failure rates with only 20% of operated rats developing the desired HF phenotype.

We describe the creation of a rat model of pressure overload induced moderate remodeling and early systolic dysfunction where signal transduction pathways involved in the initiation of the remodeling process are activated. This animal model will aid in identifying molecular targets for applying early therapeutic anti-remodeling strategies for heart failure.

Aşırı Sistolik Kalp Yetmezliği nin Aksine Aşırı Basınç Ait Orta Remodeling ve Sistolik Disfonksiyon Bir Rat Modeli

In response to an injury, such as myocardial infarction, prolonged hypertension or a cardiotoxic agent, the heart initially adapts through the activation ...