이 연구는 (심장 혈관 연구, db 위한 독일 센터), DZHK에 의해 지원 되었다 EKFS (Else Kröner Fresenius 재단, F.H.), 고 (독일 교육 및 연구), BMBF BIH Charité 임상 과학자 프로그램 투자에 의해 의해 Charité-Universitätsmedizin 베를린, 베를린 건강 연구소 (F.H.).

<ol>
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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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저자는 공개 없다.

여기, 우리는 먼저 메츠 관련 HFpEF 보여 주는 심 방 개장22표시의 쥐 모델에서 단일 셀 Acm의 격리에 대 한 프로토콜을 설명 합니다. 절차는 수술 준비로 점점 어려운 대동맥의 cannulation 과도 한 지방 조직을 만들 수 있는 유일 하 게 도전. 표 2 주소 격리 절차의 가장 일반적인 문제 문제 해결 가이드 제공.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="...

메츠 당뇨병 mellitus 타입-2와 심혈 관 질환 위험 요인의 클러스터를 설명 하 고 포함 하는 증가 동맥 혈압, dyslipidemia (트리 글리세라이드를 제기 하 고 고밀도 지 단백 콜레스테롤을 낮 췄 다), 금식 증가 포도 당, 그리고 중앙 비만1. 메츠의 전세계 보급 되며 지속적으로 상승2, 25-30%로 추정 된다. HFpEF은 메츠와 자주 관련 된 이기종 임상 증후군. HFpEF 및 그것의 이전 단계 (즉, 고혈압 성 심장 질환) 중 심장 개장은 또한 동반 atria3을 개장 하는. 감소 된 수축 성 기능 및 구조적인 변화는 왼쪽된 아 트리 움의 사망률 증가, 심 방 세 동, 및 새로운 발병 심장 마비4와 연결 되었습니다. 개장 하는 심 방 이온 채널 기능, 캘리포니아2 + 항상성, 심 방 구조, 섬유 아 세포 활성화 및 조직 섬유 증5에 변화에 의해 특징입니다. 왼쪽에 메츠 관련 HFpEF의 기본 병 적인 메커니즘 아직도 제대로 이해 하 고 더 깊이 있는 조사를 필요로 개장 하는 심 방. 동물 모델은 유용한 도구가 될 고 심 방 cardiomyopathies6,7,,89의 분야에서 많은 발전을 입증 했다.
격리 된 단일 셀 cardiomyocytes와 연구 자주 확증 vivo에서 조사 결과 보완 하는 데 사용 됩니다. 절연 및 잠재적인 후속 세포 배양, 신호 경로, 이온 채널 전류, 및 흥분-수축-커플링의 조사에 대 한 수 있습니다. 생리 적인 조건 하에서 cardiomyocytes 증식 하지 않습니다. Atrial natriuretic 요소의 transcriptional 규제 시퀀스와는 유인원 바이러스 40 큰 T 항 원 유전자 변형 쥐에서 사이의 융합 AT-110라는 첫 번째 불멸 하 게 Acm의 창조로 이끌어 냈다. AT-1 셀의 추가 개발 HL-1 셀만 순차적으로 passaged 수 없습니다 하지만 또한 저절로11계약에 상승을 했다. 그러나 그들은,, 구조와 기능 차이 덜 조직 된 열 대권 외의, hyperpolarization-활성화 안쪽으로 현재12myofibrils11, 개발의 높은 발생 등 갓 고립 된 세포에 비해 표시. 쥐와 쥐 모델의 다양 한에서 심 실 cardiomyocytes (VCM)의 절연은 잘 설립된13,14,15,16,,1718 , 19. 일반적으로, 삭제는 Langendorff 장치에 탑재 마음과 retrogradely 끼얹는다는 캘리포니아2 +-collagenases 및 프로 테아 제와 같은 소화 효소를 포함 하는 무료 버퍼. 칼슘은 다음 생리 적 조건에 단계적 방식으로 재. 그러나, Acm의 고립에 전용 프로토콜 사용할 수 있는20,21, 증가 증과 차이 압력 관련 있더라도 개장 하는 심 질환 모델에서 그들의 유용성은 제한 됩니다.
이 문서에서는, 우리는 심 방 개장 (즉, 특히 메츠 관련 HFpEF의 ZFS1 쥐 모델에 대 한)22를 나타내는 동물에서 심 방 단일 셀 cardiomyocytes의 격리에 대 한 프로토콜을 실행 했다. 기존 격리 프로토콜 최적화 하 고 제어 하 고 형태학 상으로 그리고 기능적으로 그대로 cardiomyocytes의 높은 수율으로 이어지는 심장 충 치의 intraluminal 압력 수정 간단 하 고, 주문 품 장치에 의해 보완 했다. 다음 프로토콜 단계별 가이드와 함께 연구원, 주문 품 장비, 솔루션의 목록 뿐만 아니라 포괄적인 문제 해결 가이드에 대 한 자세한 설명을 제공합니다.

<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>
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		<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>
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		<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>
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		<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>
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		<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>
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		<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>
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		<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>
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		<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

이 문서에서는, 우리는 변화 증후군 (메츠)의 쥐 모델에서 단일 셀 심 방 cardiomyocytes (Acm)의 분리에 대 한 최적화, Langendorff 기반 절차 설명-관련 보존된 방출 분수 (HFpEF)와 함께 심장 마비. 메츠 관련 HFpEF의 보급은 상승 하 고, 그리고 심 cardiomyopathies 개장 하는 심 방 및 심 방 세 동와 관련 된 관련성이 높은 임상으로 심 방 개장 사망률의 독립적인 예측 인자. 격리 된 단일 셀 cardiomyocytes와 연구 자주 확증 vivo에서 조사 결과 보완 하는 데 사용 됩니다. 순환 선박 rarefication 및 조직 간 질 성 섬유 증이이 질환의 동물 모델에서 Acm의 성공적인 단일 셀 격리에 대 한 잠재적으로 제한 요소를 포즈.
우리가 실질적으로 형태학 상으로 그리고 기능적으로 그대로 Acm의 수확량을 증가 수동으로 격리 절차 동안 심장 구멍의 intraluminal 압력을 조절 하는 장치를 사용 하 여이 문제를 해결 했습니다. 인수 세포 세포 배양 및 기능성 칼슘 이미징 (즉, 흥분-수축-커플링) 등 다양 한 실험의 다양 한에서 사용할 수 있습니다.
우리 연구원 단계별 프로토콜, 최적화 된 솔루션, 필요한 장비 및 포괄적인 문제 해결 가이드를 준비 하는 철저 한 지침의 목록 제공 한다. 절차의 초기 구현을 오히려 어려울 수도 있습니다, 그러나 성공적인 적응 메츠 관련 광범위 한 실험에 대 한 HFpEF의 쥐 모델에서 최신의 ACM 격리를 수행 하는 독자 수 있게 됩니다.

나이의 21 주에 가능한 Acm (6.1 단계에 설명 된 대로 추정), 칼슘 다시 적응 (단계 5.4-5.7), 후의 60-90% (그림 4A)이 메서드에서 ZSF-1 비만 쥐에서 분리 수 있습니다. 쥐, Acm 다르고 VCMs24,25에 비해 더 많은 이기종 표현 형 특징입니다. 그림 4B 와 보존된 막 sarcomere 구조를 기능적으로 통합 셀의 ...

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

심 방 Cardiomyocytes 보존된 방출 분수와 함께 대사 증후군 관련 심장 마비의 쥐 모델에서의 격리

여기, 우리가 보존된 방출 분수와 함께 대사 증후군 관련 심장 마비의 쥐 모델에서 단일 셀 심 방 cardiomyocytes의 격리에 대 한 최적화, Langendorff 기반 절차를 설명합니다. 심장 구멍의 intraluminal 압력의 수동 조절 기능 그대로 myocytes 흥분-수축-결합 연구에 대 한 적합 한를 구현 됩니다.

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 조회수.  Charité University Medicine. 여기, 우리가 보존된 방출 분수와 함께 대사 증후군 관련 심장 마비의 쥐 모델에서 단일 셀 심 방 cardiomyocytes의 격리에 대 한 최적화, Langendorff 기반 절차를 설명합니다. 심장 구멍의 intraluminal 압력의 수동 조절 기능 그대로 myocytes 흥분-수축-결합 연구에 대 한 적합 한를 구현 됩니다.

비디오: 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.

잠복기 모델에서 허혈성 심장 마비에 대한 유전자 전송

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

연구

의학

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.

경비원이 심방세동 세동에 대한 심방세동 부위요 폐쇄 장치를 왼쪽

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.

새로운 싱글 회의소 Implantable 심방세동 감지와 심장 박동기 : 감지의 실용 데모와 주입의 용이성

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.

고립의 방법<em&gt; 전의 VIVO</em쥐 모델에서&gt; 폐 관류 : 쥐 EVLP 프로그램 개발에서 교훈을

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.

고립 된 랫트 심장으로부터의 간질과 투과 물 수집을위한 역전 된 심장 모델

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.

절연 및 성인 쥐 Cardiomyocytes의 재배

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.

돼지에서 만성 심장 마비 모델로 tachycardia 유발 심근

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.

성인 마우스에서 심방 근세포의 격리

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.

줄기 세포 치료를위한 심근 경색 심부전의 돼지 모델 설립

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.

압력 과부하의 쥐 모델은 오버트 수축기 심부전과 반대로 적당한 리모델링 및 수축기 기능 장애를 유도했습니다.

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