D. H. Kim thanks the Department of Bioengineering at the University of Washington for the new faculty startup fund. D. H. Kim is also supported by the Perkins Coie Award for Discovery, the Wallace H. Coulter Foundation Translational Research Partnership Award, the Washington State Life Science Discovery Fund, and the American Heart Association Scientist Development Grant (13SDG14560076). J. Macadangdang and A. Jiao thank the support from the NIH Bioengineering Cardiovascular Training Grant Fellowship. &#160;Additional support for this work comes from the National Institutes of Health (NIH) grant R01HL111197 to M. Regnier.

별도의 표시가없는 한, 모든 절차는 실온 (~ 23 ° C)에서 실시하고 있습니다. 실리콘 마스터 1. 제작 <ol><li> O 2 / N 2 가스에서 100 % 에탄올, 자일 렌 및 건조와 깨끗한 실리콘 웨이퍼. </li><li> 0.3 ~ 0.5 μm의 두께 필름을 생산하기 위해 2,000-4,000 RPM의 회전 속도로 스핀 코터 실리콘 웨이퍼를 놓습니다. </li><li> 포토 리소그래피 시스템을 사용하여 패턴 정확한 치수를 갖는 포토 레지스트 막을 </li><li> 완전 포토 레지스트 개발 솔루션의 적절한 볼륨의 패턴 화 된 포토 레지스트 코팅 된 실리콘 웨이퍼를 체험. </li><li> 탈 이온수로 개발 된 포토 레지스트 코팅 된 실리콘 웨이퍼를 씻어. </li><li> 근처 수직 측벽, 깊은 반응성 이온 식각 에칭 시스템을 사용하여 노출 된 실리콘과 함께 서브 마이크론 치수 능선 어레이를 형성한다. </li><li> 플라즈마 애셔 시스템에서 실리콘 웨이퍼를 배치하여 남은 포토 레지스트를 제거한다. </li><li> 승 실리콘을 잘라이후의 복제 성형에 대한 적절한 크기의 실리콘 마스터에 다이아몬드 팁 커터 AFERS. </li></ol> 실리콘 마스터에서 PUA 몰드 2. 제작 주 : 나노 제조를위한 실리콘 마스터에 추가 할 양이 우레탄 아크릴 레이트 (PUA) 용액의 점도뿐만 아니라 복제 될 나노 패턴 마스터의 면적에 따라 달라질 것이다. <ol><li> 100 % 에탄올 또는 O 2 / N 2 가스에서 자일 렌 및 건조와 깨끗한 실리콘 마스터면. </li><li> 페트리 접시에 실리콘 마스터 패턴 측면을 놓습니다. </li><li> 2cm X 2cm 표면 패턴, 패턴의 표면에 PUA의 피펫 40 μL와 실리콘 마스터하십시오. </li><li> 4cm 분배 PUA에 대한 X 4cm 투명한 폴리 에스테르 (PET) 필름의 시트를 놓습니다. </li><li> 아래로 PET 시트에 눌러 수 있도록 (예 : 카드 등) 롤러 또는 평면 가장자리가 표면을 사용하여 패턴면에서 시트 아래에 PUA 확산 전체패턴 PUA 프리폴리머에 의해 덮여있다. </li><li> 50 초 동안 약 10cm 20 와트 (115 V) 자외선 (λ = 365 nm의) 아래의 실리콘 마스터, 예비 및 PET를 놓습니다. 효과가, 자외선 파장은 310-400 nm의 사이에 어디 에나있을 수 있습니다. 빛의 세기는 상기 기판의 표면에서 10 ~ 15 mW의 / cm 2이다. </li><li> 경화 후, 집게 천천히 PET 필름을 제거합니다. PUA는 실리콘 마스터 나노 패턴의 음과 PET 필름에 부착해야한다. </li><li> 사용하기 전에 최소 12 시간 동안 UV에서 PUA / PET의 나노 패턴을 치료. 과다 노출은 문제가되지 않습니다. </li><li> 실리콘 마스터를 청소하려면, PUA의 추가없이 마스터 위에 PET의 또 다른 필름을 배치하고 50 초 동안 자외선 (λ = 365 nm의)에 노출 및 PET 필름을 제거합니다. 이것은 미 반응 모노머를 제거합니다. </li><li> O 2 / N 2 가스에서 100 % 에탄올, 자일 렌 및 건조와 실리콘 마스터를 씻어. </li></ol> 3A. 나노 패터닝 폴리 우레탄 폴리머 </P&gt; <ol><li> 10 분간 오존 처리 챔버 내에 배치하여 25mm 지름의 원형 유리 슬라이드를 준비한다. </li><li> 쉬운 취급을위한 작은 PDMS 블록에 오존 처리 된 유리 슬라이드를 놓습니다. </li><li> 유리 슬라이드에 페인트 브러시와 표면 접착 촉진제의 얇은 층을 적용하십시오. 30 분 동안 공기 건조 유리 슬라이드. </li><li> 프린터 종이에 유리 슬라이드를 놓습니다. </li><li> 유리 슬라이드의 중심에 사전 폴리머 (NOA 76) 폴리 우레탄 10 μL (PU)을 분배 삭제합니다. 기포가 첨가 한 후 존재하지 않는 확인하십시오. </li><li> 장소 PUA 금형, 패턴 유리 슬라이드에, 얼굴을 아래로. 균일 PUA 몰드 따라 실린더 고무 롤러를 회전함에 따라 유리 슬라이드의 표면에 걸쳐 PU를 분산. 프린터 용지 폴리머 오버 플로우를 흡수 할 것이다. </li><li> 와트 UV 램프. PU의 중합 시간은 UV 소스의 힘에 의존한다. </li><li> UV 광원으로부터 샘플을 제거하고주의 깊게 PU 코팅 된 유리 슬라이드에서 PUA 몰드 껍질. PU의 중합 C로 간주됩니다omplete 완전히 떨어져 샘플에서 PUA 몰드 껍질과 PU 유리 슬라이드 무지개 빛깔의 외관이 때. </li><li> 한 달에 스토리지를위한 데시 케이 터에서 완성 샘플을 놓습니다. </li></ol> (b). 나노 패터닝 폴리 (락 타이드 - 코 - 글리콜 라이드) 히드로 겔 <ol><li> 적극적으로 10:1의 비율로 실리콘 엘라스토머 기지와 실리콘 엘라스토머 경화제를 혼합하여 평면 PDMS 몰드를 만듭니다. </li><li> (PDMS 5 ㎜ 두께가되도록, 즉) PDMS 전구체 접시의 가장자리를 5mm까지 도달하도록 혼합 PDMS가 페트리 디쉬에 액을 전구체 붓는다. </li><li> 가스를 제거하는 1 시간 동안 건조기에 페트리 접시와 PDMS 전구체를 놓습니다. </li><li> 치료하기 위해 적어도 2 시간 동안 65 ° C의 오븐에 페트리 접시와 PDMS 전구체를 이동합니다. </li><li> PDMS는 치료 후 3cm × 3 인치의 정사각형 섹션으로 평면 PDMS를 잘라 면도기를 사용합니다. 이러한 패터닝 공정에서 나중에 사용 평 PDMS 몰드 것이다. </li><li> PL 청결한 지름 25mm 원형 유리 슬라이드물을 초음파 처리기에 30 분 동안 이소 프로필 알콜로 acing. </li><li> O 2 / N 2 가스 하에서 드라이 세정 된 유리 슬라이드. </li><li> 드롭 PLGA 용액 100 μl를 분배 유리 슬라이드에 (15 % w 클로로포름 PLGA V /). </li><li> 용매를 흡수하고 평평한 PLGA 표면을 얻기 위해 분배 된 PLGA 위에 평평한 PDMS 몰드를 놓습니다. 5 분 동안 PDMS 위에 200g의 중량을 배치하여 가벼운 압력 (~ 10 kPa의)을 적용한다. </li><li> 서서히 평 PDMS 몰드를 박리하고 잔류 용매를 제거하고 PLGA와 커버 글라스 사이의 밀착성을 증가시키기 위해 5 분 동안 예열 플레이트 (120 ℃)​​에 커버 글라스를 배치. </li><li> 플랫 PLGA 위에 NP PUA 몰드를 놓고 PUA 몰드의 상단에 1 kg 무게를 배치하여 일정한 압력 (~ 100 kPa의)과 열 (120 ° C)를 적용 15 분 동안 가열 접시에있다. </li><li> PLGA 커버 슬라이드에서 무게를 제거하고 하층은 실온으로 냉각시킨다. 하층이 될 때까지 PUA 몰드를 제거하지 마십시오냉각. </li><li> 하층가 충분히 냉각 한 후, 조심스럽게 NP PLGA의 기질을 드러내는, NP PUA 몰드를 벗겨. NP PLGA의 토대는 무지개 빛깔의 모습이 있어야합니다. </li><li> 한 달에 스토리지를위한 데시 케이 터에서 완성 샘플을 놓습니다. </li></ol> 4. 셀 시드와 문화 참고 :이 프로토콜은 신생아 쥐의 심실 근육 세포 (NRVMs)과 H7 인간 배아 줄기 세포 유래 심근 (hESC의-CMS)하지만 다른 세포 소스의 문화를 설명 사용할 수있다. <ol><li> 35mm 조직 문화 폴리스티렌 접시에 ANFS의 커버 슬립 (PU 또는 PLGA)를 연결합니다. 접시의 바닥에 조심스럽게 놀 랜드 광학 접착제 (NOA83H)의 피펫 20 μL는 NOA의 상단에 ANFS coverslip에 배치합니다. 접착제가 퍼져 전체 coverslip에 바닥을 커버 할 수 있습니다. 10 분 동안 UV에 요리를 노출하여 NOA 치료. </li><li> 회, 5 분 동안 70 % 에탄올 수용액 2 ㎖로 세정하여 ANFS 살균. 전자를 제거흡인 thanol. 허용 ANFS 생물 안전 캐비닛의 UV 살균 램프 (λ = 200-290 nm의)에서 ~ 1 시간 동안 완전히 건조 공기. </li><li> 세포 접착 밤새 피브로넥틴에 ANFS 코팅에 의해 강화된다. 5 ㎍ / ml로 DI 물에 피브로넥틴을 희석. 접시에 피브로넥틴 솔루션의 피펫 2 ㎖. 37 ° C에서 인큐베이터에 배치하고 5 % CO 2 밤새 (최소 6 시간). </li><li> 이전 프로토콜 22에 따라 NRVMs, hESC의 CMS가, 또는 관심의 다른 심장 세포를 얻습니다. </li><li> 펠릿 세포에 3 분 동안 1,000 rpm으로 원심 분리기 세포 샘플. </li><li> 흡인에 의해 조심스럽게 뜨는을 제거합니다. 펠렛을 방해하지 않도록하십시오. </li><li> 4.6 × 106 세포 / ml의 농도로 적절한 배양 배지에서 세포를 재현 탁. </li><li> 조심스럽게 살균 ANFS에 세포 현탁액 200 μl를 피펫. 확인 세포 현탁액은 커버 슬립에 남아합니다. </li><li> 37 ° C와 5 % 배양기에서 세포를 배치CO 세포가 ANFS에 부착 할 수 있도록 4 시간 2. </li><li> 동일한 조건에서 배양기에서 세포를 접시 및 교체 따뜻한 배지 2 ㎖를 추가합니다. </li><li> 24 시간 후, 미디어를 제거하고 여분의 세포를 제거하기 위해 두 번 DPBS 2 ㎖로 씻는다. </li><li> 동일한 조건에서 배양기에서 세포를 접시 및 교체 따뜻한 배지 2 ㎖를 추가합니다. 문화 세포가 합류합니다. 매일 미디어를 교체합니다. </li></ol>

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Authors have nothing to disclose.

기능적으로 성숙한 심장 조직은 생체 내에서와 심장 조직 공학 생체 응용 프로그램에서 모두 부족하다. 여기에 설명 CFL의 나노 제조 방법은 셀룰러 정렬을 달성 인해 시스템의 확장 성으로 거시적 조직 기능을 좌우위한 견고한 기술이다. 큰 영역은 쉽게 패턴과 세포 배양에 사용할 수 있습니다. 매크로 셀 정렬은 생체 모방, 그것은 심근 (38)의 기계 및 전기 두 속성에 영향을 ...

심혈관 질환은 세계에서 사망률 및 사망의 주요 원인이며, 이미 변형 된 세계 보건 시스템 1,17에 무거운 사회 경제적 부담을 제시한다. (1) 허혈성 질환이나 심근 병증 후 손상된 심근을 재생하거나 (2) 생체 외 약물 검사 또는 질병 모델링을위한 마음의 고 충실도 모델을 만드는 : 심장 조직 공학은 두 가지 목표를 가지고 있습니다. 심장은 몸에 혈액을 공급하기 위해 지속적으로 작동해야합니다 복잡한 기관이다. 심근 및 지원 조직의 밀집 층 구조는 심장 벽 (18, 19)에 걸쳐 나선형 패턴으로 배치되어있다. 심장은 전기 기계적 효율적으로 본체 (21)에 혈액을 배출 할 수있는 고도의 조정 방식으로 20에 연결된다. 자연의 아야 신뢰성 시험 관내에서 효과적으로 요약 될 수 있기 전에 여러 주요 장애물은, 그러나, 해결되어야 남아있다.강력한 심근 분화 방법 (22)을 개발하기 위해 계속하지만, 첫째, hPSC-CMS는 여전히 다소 미숙 표현형을 나타낸다. 자신의 전기 특성과 형태에 가장 가까운 태아의 수준 23 일치. 전통 문화의 조건에서 보관하는 경우 둘째, 줄기 세포 유래 및 기본 심근 모두 기본, 조직과 같은 구조로 조립하는 데 실패합니다. 오히려 세포가 무작위로 지향되고 성숙한 심근 (24)의 줄무늬 막대 모양의 외관을 지원하지 않는 경우가 있습니다. 세포가 상호 작용과 세포 외 기질 (ECM) 환경은 많은 세포 과정 11,13,25에 중요한 역할을한다. ECM은 크게 6,26 세포의 구조 및 기능에 영향을 미치는 복합, 잘 정의 된 분자 및 지형 큐들로 구성된다. 심장 내에서 세포 정렬 밀접 기본 나노 미터 규모의 ECM 섬유 2를 다음과 같습니다. 이러한 nanotopograph의 영향세포와 조직의 기능에 iCal의 단서는, 그러나, 지금까지 완전히 이해에 있습니다. 나노 미터 크기의 세포 - 생체 적합 물질의 상호 작용의 예비 연구 (27) 세포 신호 전달, 접착 28-30, 성장 31, 32, 33 및 차별화를위한 서브 마이크론 지형 신호의 잠재적 인 중요성과 효과를 나타냅니다. 그러나, 재현과 확장 성 나노 제조 기판 개발에 어려움에, 이러한 연구는 생체 내 ECM 환경에서 복잡한 멀티 스케일 휴대 효과를 재현 할 수 있습니다. 이 프로토콜에서, 네이티브 심장 ECM 섬유 배향을 흉내 낸 세포 배양 지지체를 생성하는 간단하고 비용 효과적인 나노 제조 기술은 심근 세포 - 생체 물질의 상호 작용의 신규 한 조사 광범위한 있도록 설명한다. 심근은 나노 ECM 환경과 상호 작용하는 방식을 이해하는 것은 더 가깝게 모방 기본 조직의 기능에 휴대 동작을 제어 할 수있는 능력을 허용 할 수기. 또한, 세포 단일 층은 3 차원 구조에 비해 단순화 된 실험 시스템입니다 만, 여전히 통찰력 조사와 기능 검사 2,34-36 복잡한 다세포 동작을 나타냅니다. 마지막으로, 지지체 (37)는 재생 목적을 위해 심장에 이식 할 때 셀룰러 그래프트 기능을 향상시키기 위해 사용될 수있다.

<table border="0" cellpadding="0" cellspacing="0" width="352">
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	<tbody>
		<tr height="43" style="height:32.25pt">
			<td class="xl63" height="43" style="height:32.25pt;width:66pt" width="88">Name</td>
			<td class="xl63" style="width:66pt" width="88">Company</td>
			<td class="xl63" style="width:66pt" width="88">Catalogue number</td>
			<td class="xl64" style="width:66pt" width="88">Comments (optional)</td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Fibronectin</td>
			<td class="xl65" style="width:66pt" width="88">BD Biosciences</td>
			<td class="xl65" style="width:66pt" width="88">354008</td>
			<td class="xl66" style="width:66pt" width="88"></td>
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		<tr height="64" style="height:48.0pt">
			<td class="xl65" height="64" style="height:48.0pt;width:66pt" width="88">NOA 76</td>
			<td class="xl65" style="width:66pt" width="88">Norland Products, Inc.</td>
			<td class="xl65" style="width:66pt" width="88">7606B</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="106" style="height:79.5pt">
			<td class="xl65" height="106" style="height:79.5pt;width:66pt" width="88">Surface Adhesion Promotor (Glass Primer)</td>
			<td class="xl65" style="width:66pt" width="88">Minuta Tech</td>
			<td class="xl65" style="width:66pt" width="88"></td>
			<td class="xl66" style="width:66pt" width="88"></td>
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		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">PUA</td>
			<td class="xl65" style="width:66pt" width="88">Minuta Tech</td>
			<td class="xl65" style="width:66pt" width="88">MINS-311RM</td>
			<td class="xl66" style="width:66pt" width="88"></td>
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		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Soft Rubber Roller</td>
			<td class="xl65" style="width:66pt" width="88">Speedball</td>
			<td class="xl65" style="width:66pt" width="88"></td>
			<td class="xl66" style="width:66pt" width="88"></td>
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		<tr height="64" style="height:48.0pt">
			<td class="xl65" height="64" style="height:48.0pt;width:66pt" width="88">Silicon Wafers</td>
			<td class="xl65" style="width:66pt" width="88">NOVA Electronic Materials</td>
			<td class="xl65" style="width:66pt" width="88">FA01-9900</td>
			<td class="xl66" style="width:66pt" width="88"></td>
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		<tr height="22" style="height:16.5pt">
			<td class="xl65" height="22" style="height:16.5pt;width:66pt" width="88">Photoresist</td>
			<td class="xl65" style="width:66pt" width="88">Shipley</td>
			<td class="xl65" style="width:66pt" width="88">SPRT510</td>
			<td class="xl66" style="width:66pt" width="88"></td>
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		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Photoresist Developer</td>
			<td class="xl65" style="width:66pt" width="88">Shipley</td>
			<td class="xl65" style="width:66pt" width="88">MF320</td>
			<td class="xl66" style="width:66pt" width="88"></td>
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		<tr height="85" style="height:63.75pt">
			<td class="xl65" height="85" style="height:63.75pt;width:66pt" width="88">Electron-Beam Lithography System</td>
			<td class="xl65" style="width:66pt" width="88">JEOL</td>
			<td class="xl65" style="width:66pt" width="88">JBX-9300FS</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="64" style="height:48.0pt">
			<td class="xl65" height="64" style="height:48.0pt;width:66pt" width="88">Etching System</td>
			<td class="xl65" style="width:66pt" width="88">Surface Technology Systems</td>
			<td class="xl65" style="width:66pt" width="88">NP10 8UJ</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="64" style="height:48.0pt">
			<td class="xl65" height="64" style="height:48.0pt;width:66pt" width="88">Plasma Asher System</td>
			<td class="xl65" style="width:66pt" width="88">BMR Technology Co.</td>
			<td class="xl65" style="width:66pt" width="88">DSF-200</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Ozone Cure System</td>
			<td class="xl65" style="width:66pt" width="88">Minuta Tech</td>
			<td class="xl65" style="width:66pt" width="88">MT-UV-O- 08</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Fusion Cure System</td>
			<td class="xl65" style="width:66pt" width="88">Minuta Tech</td>
			<td class="xl65" style="width:66pt" width="88">MT-UV-A 11</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="64" style="height:48.0pt">
			<td class="xl65" height="64" style="height:48.0pt;width:66pt" width="88">NOA 83H</td>
			<td class="xl65" style="width:66pt" width="88">Norland Products, Inc.</td>
			<td class="xl65" style="width:66pt" width="88">8301</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Spin Coater</td>
			<td class="xl65" style="width:66pt" width="88">Laurel Technology</td>
			<td class="xl65" style="width:66pt" width="88">WS-400-6NPP</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Skyrol PET Film</td>
			<td class="xl65" style="width:66pt" width="88">SKC Co., Ltd.</td>
			<td class="xl65" style="width:66pt" width="88">23038-59-9</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">25mm Glass Slides</td>
			<td class="xl65" style="width:66pt" width="88">Corning</td>
			<td class="xl65" style="width:66pt" width="88">2948</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="85" style="height:63.75pt">
			<td class="xl65" height="85" style="height:63.75pt;width:66pt" width="88">Sylgard 184 Silicone Elastomer Kit</td>
			<td class="xl65" style="width:66pt" width="88">Dow Corning</td>
			<td class="xl67" style="width:66pt" width="88">6/5/2553</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="64" style="height:48.0pt">
			<td class="xl65" height="64" style="height:48.0pt;width:66pt" width="88">Poly(D,L-lactide-co-glycolide)</td>
			<td class="xl65" style="width:66pt" width="88">Sigma-Aldrich</td>
			<td class="xl65" style="width:66pt" width="88">P2191-1G</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Chloroform</td>
			<td class="xl65" style="width:66pt" width="88">Sigma-Aldrich</td>
			<td class="xl65" style="width:66pt" width="88">372978-1L</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">500g Weights</td>
			<td class="xl65" style="width:66pt" width="88">Global Insustrial</td>
			<td class="xl65" style="width:66pt" width="88">T9FB503120</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="43" style="height:32.25pt">
			<td class="xl65" height="43" style="height:32.25pt;width:66pt" width="88">Isopropyl Alcohol</td>
			<td class="xl65" style="width:66pt" width="88">EMD Millipore</td>
			<td class="xl65" style="width:66pt" width="88">PX1835-2</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="22" style="height:16.5pt">
			<td class="xl65" height="22" style="height:16.5pt;width:66pt" width="88">Hot Plate</td>
			<td class="xl65" style="width:66pt" width="88">Corning</td>
			<td class="xl65" style="width:66pt" width="88">PC-420D</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
		<tr height="22" style="height:16.5pt">
			<td class="xl65" height="22" style="height:16.5pt;width:66pt" width="88">Sonicator</td>
			<td class="xl65" style="width:66pt" width="88">Branson</td>
			<td class="xl65" style="width:66pt" width="88">B2510MTH</td>
			<td class="xl66" style="width:66pt" width="88"></td>
		</tr>
	</tbody>
</table>

capillary force lithography for cardiac tissue engineering

심혈관 질환은 전세계 사망 1의 주요 원인이 남아있다. 심장 조직 공학 심장 재생뿐만 아니라 체외 심사 분석을위한 기능적인 조직 개발의 목표와 혁신적인 의료 발견을 제공하기 위해 많은 약속을 보유하고 있습니다. 그러나, 심장 조직의 충실도가 높은 모델을 만들 수있는 능력이 어려운 입증했다. 심장의 세포 외 기질 (ECM)는에서 나노 미터 규모의 2 마이크로에 이르기까지 모두 생화학 및 생 역학적 인 신호로 구성된 복잡한 구조입니다. 기계적 하중 조건과 세포 - ECM 상호 작용 지역은 최근 심장 조직 공학 3-5의 중요한 구성 요소로 인식되고있다. 심장 ECM의 많은 부분이 크게 조직 구조 및 전기 기계 결합이 영향을 나노 스케일 직경 정렬 콜라겐 섬유로 구성된다. 불행하게도, 몇 가지 방법이 근래전자 나노 미터 크기까지 ECM 섬유의 조직을 모방 할 수있게되었습니다. 나노 제조 기술의 최근 발전은, 그러나, 심장 6-9에서 ECM의 생체 구조 및 기판 강성 큐들을 모방 확장 발판의 설계 및 제조를 사용할 수있다. 여기에서 우리는의 발전을 제시이 재현성, 경제적, 및 생체 적합성 중합체, 폴리 (락 티드 - 코 - 글리콜 라이드) (PLGA) (8) 및 폴리 우레탄 (PU) 계 중합체를 사용하여 심장 세포의 기능적 정렬을위한 확장 가능한 나노 패터닝 공정. 이러한 이방를 nanofabricated 하층 (ANFS는) 잘 조직 된, 정렬 조직의 기본 ECM을 모방하고 세포 형태 및 기능을 10 ~ 14에 나노 형상의 역할을 조사 할 수 있습니다. 주형으로 나노 패턴 (NP) 실리콘 마스터를 사용하여, 폴리 우레탄 아크릴 레이트 (PUA) 금형을 제작한다. 이 PUA 몰드는 다음 파에 사용된다각각 UV-지원 또는 용매 매개 모세관 력 리소그래피 (CFL), 15, 16을 통해 PU 또는 PLGA 하이드로 겔을 ttern. 간단히, PU 또는 PLGA 예비 중합체는 유리 커버 슬립하고 PUA 금형이 상부에 배치되어 상으로 분배 된 드롭이다. UV 보조 CFL 들어, PU는 후 경화 UV 방사선 (λ = 250-400 NM)에 노출된다. 용매 매개 CFL의 경우, PLGA는 열 (120 ° C) 및 압력 (100 kPa의)를 사용하여 양각되어있다. 경화 후, PUA 몰드는 세포 배양을위한 ANFS 남기고 박리된다. 이러한 신생아 래트 심실 근세포뿐만 아니라 인간 다 능성 줄기 세포 유래 심근 같은 일차 전지는, ANFS 2에 유지 될 수있다.

도 1은이 제조 방법에 대한 제조 공정의 개략도이다. 때문에 나노 지형에 의한 빛의 회절, 나노 패터닝은 ANFS에 무지개 빛깔의 표면에 발생한다. 2는 800 nm의 능선과 홈에 잘 패턴 25mm NP-PU coverslip에 (그림 2A)에서이 무지개 빛깔의 표면을 묘사 그림 폭 (그림 2B). ANF​​S의 무지개 빛깔의 외관은 능선과 홈의 폭에 따라 다소 차이가 있습?...

Watch this Scientific Journal Video about Capillary Force Lithography for Cardiac Tissue Engineering at JoVE.com

Capillary Force Lithography for Cardiac Tissue Engineering

In this protocol, we demonstrate the fabrication of biomimetic cardiac cell culture substrata made from two distinct polymeric materials using capillary force lithography. The described methods provide a scalable, cost-effective technique to engineer the structure and function of macroscopic cardiac tissues for in vitro and in vivo applications.

심장 조직 공학을위한 모세관 력 리소그래피

Cardiovascular disease remains the leading cause of death worldwide1. Cardiac tissue engineering holds much promise to deliver groundbreaking medical ...

capillary-force-lithography-for-cardiac-tissue-engineering

Research

JoVE Journal

Bioengineering

12.3K 조회수.  University of Washington. In this protocol, we demonstrate the fabrication of biomimetic cardiac cell culture substrata made from two distinct polymeric materials using capillary force lithography. The described methods provide a scalable, cost-effective technique to engineer the structure and function of macroscopic cardiac tissues for in vitro and in vivo applications.

비디오: Capillary Force Lithography for Cardiac Tissue Engineering

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications. Briefly, decellularized tissue is lyophilized, milled, enzymatically digested, and then brought to physiological pH. The lyophilization removes all water content from the tissue, resulting in dry ECM that can be ground into a fine powder with a small mill. After milling, the ECM powder is digested with pepsin to form an injectable matrix. After adjustment to pH 7.4, the liquid matrix material can be injected into the myocardium. Results of previous characterization assays have shown that matrix gels produced from decellularized pericardial and myocardial tissue retain native ECM components, including diverse proteins, peptides and glycosaminoglycans. Given the use of this material for tissue engineering, in vivo characterization is especially useful; here, a method for performing an intramural injection into the left ventricular (LV) free wall is presented as a means of analyzing the host response to the matrix gel in a small animal model. Access to the chest cavity is gained through the diaphragm and the injection is made slightly above the apex in the LV free wall. The biologically derived scaffold can be visualized by biotin-labeling before injection and then staining tissue sections with a horse radish peroxidase-conjugated neutravidin and visualizing via diaminobenzidine (DAB) staining. Analysis of the injection region can also be done with histological and immunohistochemical staining. In this way, the previously examined pericardial and myocardial matrix gels were shown to form fibrous, porous networks and promote vessel formation within the injection region.

Methods for preparing an injectable matrix gel from decellularized tissue and injecting it into rat myocardium in vivo are described.

심근 조직 공학을위한 생물학 유래 주사용 재료의 제조

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications.  ...

연구

생체공학

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and increasing obesity1. Currently, bypass surgery using autologous vessels, allografts, and synthetic grafts are known as a commonly used for arterial substitutes2. However, these grafts have limited applications when an inner diameter of arteries is less than 6 mm due to low availability, thrombotic complications, compliance mismatch, and late intimal hyperplasia3,4. To overcome these limitations, tissue engineering has been successfully applied as a promising alternative to develop small-diameter arterial constructs that are nonthrombogenic, robust, and compliant. Several previous studies have developed small-diameter arterial constructs with tri-lamellar structure, excellent mechanical properties and burst pressure comparable to native arteries5,6. While high tensile strength and burst pressure by increasing collagen production from a rigid material or cell sheet scaffold, these constructs still had low elastin production and compliance, which is a major problem to cause graft failure after implantation. Considering these issues, we hypothesized that an elastometric biomaterial combined with mechanical conditioning would provide elasticity and conduct mechanical signals more efficiently to vascular cells, which increase extracellular matrix production and support cellular orientation.
The objective of this report is to introduce a fabrication technique of porous tubular scaffolds and a dynamic mechanical conditioning for applying them to arterial tissue engineering. We used a biodegradable elastomer, poly (glycerol sebacate) (PGS)7 for fabricating porous tubular scaffolds from the salt fusion method. Adult primary baboon smooth muscle cells (SMCs) were seeded on the lumen of scaffolds, which cultured in our designed pulsatile flow bioreactor for 3 weeks. PGS scaffolds had consistent thickness and randomly distributed macro- and micro-pores. Mechanical conditioning from pulsatile flow bioreactor supported SMC orientation and enhanced ECM production in scaffolds. These results suggest that elastomeric scaffolds and mechanical conditioning of bioreactor culture may be a promising method for arterial tissue engineering.

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

탄성 PGS는 아티 조직 공학 공사장 공중 발판

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

Plasma Lithography Surface Patterning for Creation of Cell Networks

Systematic manipulation of a cell microenvironment with micro- and nanoscale resolution is often required for deciphering various cellular and molecular phenomena. To address this requirement, we have developed a plasma lithography technique to manipulate the cellular microenvironment by creating a patterned surface with feature sizes ranging from 100 nm to millimeters. The goal of this technique is to be able to study, in a controlled way, the behaviors of individual cells as well as groups of cells and their interactions.
This plasma lithography method is based on selective modification of the surface chemistry on a substrate by means of shielding the contact of low-temperature plasma with a physical mold. This selective shielding leaves a chemical pattern which can guide cell attachment and movement. This pattern, or surface template, can then be used to create networks of cells whose structure can mimic that found in nature and produces a controllable environment for experimental investigations. The technique is well suited to studying biological phenomenon as it produces stable surface patterns on transparent polymeric substrates in a biocompatible manner. The surface patterns last for weeks to months and can thus guide interaction with cells for long time periods which facilitates the study of long-term cellular processes, such as differentiation and adaption. The modification to the surface is primarily chemical in nature and thus does not introduce topographical or physical interference for interpretation of results. It also does not involve any harsh or toxic substances to achieve patterning and is compatible for tissue culture. Furthermore, it can be applied to modify various types of polymeric substrates, which due to the ability to tune their properties are ideal for and are widely used in biological applications. The resolution achievable is also beneficial, as isolation of specific processes such as migration, adhesion, or binding allows for discrete, clear observations at the single to multicell level.
This method has been employed to form diverse networks of different cell types for investigations involving migration, signaling, tissue formation, and the behavior and interactions of neurons arraigned in a network.

A versatile plasma lithography technique has been developed to generate stable surface patterns for guiding cellular attachment. This technique can be applied to create cell networks including those that mimic natural tissues and has been used for studying several, distinct cell types.

세포 네트워크의 창조를위한 플라즈마 리소그래피 표면 Patterning

Systematic manipulation of a cell microenvironment with micro- and nanoscale resolution is often required for deciphering various cellular and molecular ...

Encapsulation of Cardiomyocytes in a Fibrin Hydrogel for Cardiac Tissue Engineering

Culturing cells in a three dimensional hydrogel environment is an important technique for developing constructs for tissue engineering as well as studying cellular responses under various culture conditions in vitro. The three dimensional environment more closely mimics what the cells observe in vivo due to the application of mechanical and chemical stimuli in all dimensions 1. Three-dimensional hydrogels can either be made from synthetic polymers such as PEG-DA 2 and PLGA 3 or a number of naturally occurring proteins such as collagen 4, hyaluronic acid 5 or fibrin 6,7. Hydrogels created from fibrin, a naturally occurring blood clotting protein, can polymerize to form a mesh that is part of the body's natural wound healing processes 8. Fibrin is cell-degradable and potentially autologous 9, making it an ideal temporary scaffold for tissue engineering.
Here we describe in detail the isolation of neonatal cardiomyocytes from three day old rat pups and the preparation of the cells for encapsulation in fibrin hydrogel constructs for tissue engineering. Neonatal myocytes are a common cell source used for in vitro studies in cardiac tissue formation and engineering 4. Fibrin gel is created by mixing fibrinogen with the enzyme thrombin. Thrombin cleaves fibrinopeptides FpA and FpB from fibrinogen, revealing binding sites that interact with other monomers 10. These interactions cause the monomers to self-assemble into fibers that form the hydrogel mesh. Because the timing of this enzymatic reaction can be adjusted by altering the ratio of thrombin to fibrinogen, or the ratio of calcium to thrombin, one can injection mold constructs with a number of different geometries 11,12. Further we can generate alignment of the resulting tissue by how we constrain the gel during culture 13.
After culturing the engineered cardiac tissue constructs for two weeks under static conditions, the cardiac cells have begun to remodel the construct and can generate a contraction force under electrical pacing conditions 6. As part of this protocol, we also describe methods for analyzing the tissue engineered myocardium after the culture period including functional analysis of the active force generated by the cardiac muscle construct upon electrical stimulation, as well as methods for determining final cell viability (Live-Dead assay) and immunohistological staining to examine the expression and morphology of typical proteins important for contraction (Myosin Heavy Chain or MHC) and cellular coupling (Connexin 43 or Cx43) between myocytes.

We describe the isolation of neonatal cardiomyocytes and the preparation of the cells for encapsulation in fibrin hydrogel constructs for tissue engineering. We describe methods for analyzing the tissue engineered myocardium after the culture period including active force generated upon electrical stimulation and cell viability and immunohistological staining.

심장 조직 공학을위한 섬유소 히드로겔에 Cardiomyocytes의 캡슐

Culturing cells in a three dimensional hydrogel environment is an important technique for developing constructs for tissue engineering as well as studying ...

Postproduction Processing of Electrospun Fibres for Tissue Engineering

Electrospinning is a commonly used and versatile method to produce scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo undergo biaxial distension to varying extents such as skin, bladder, pelvic floor and even the hard palate as children grow. In producing scaffolds for these purposes there is a need to develop scaffolds of appropriate biomechanical properties (whether achieved without or with cells) and which are sterile for clinical use. The focus of this paper is not how to establish basic electrospinning parameters (as there is extensive literature on electrospinning) but on how to modify spun scaffolds post production to make them fit for tissue engineering purposes - here thickness, mechanical properties and sterilisation (required for clinical use) are considered and we also describe how cells can be cultured on scaffolds and subjected to biaxial strain to condition them for specific applications.
Electrospinning tends to produce thin sheets; as the electrospinning collector becomes coated with insulating fibres it becomes a poor conductor such that fibres no longer deposit on it. Hence we describe approaches to produce thicker structures by heat or vapour annealing increasing the strength of scaffolds but not necessarily the elasticity. Sequential spinning of scaffolds of different polymers to achieve complex scaffolds is also described. Sterilisation methodologies can adversely affect strength and elasticity of scaffolds. We compare three methods for their effects on the biomechanical properties on electrospun scaffolds of poly lactic-co-glycolic acid (PLGA).
Imaging of cells on scaffolds and assessment of production of extracellular matrix (ECM) proteins by cells on scaffolds is described. Culturing cells on scaffolds in vitro can improve scaffold strength and elasticity but the tissue engineering literature shows that cells often fail to produce appropriate ECM when cultured under static conditions. There are few commercial systems available that allow one to culture cells on scaffolds under dynamic conditioning regimes - one example is the Bose Electroforce 3100 which can be used to exert a conditioning programme on cells in scaffolds held using mechanical grips within a media filled chamber.4 An approach to a budget cell culture bioreactor for controlled distortion in 2 dimensions is described. We show that cells can be induced to produce elastin under these conditions. Finally assessment of the biomechanical properties of processed scaffolds cultured with or without cells is described.

Electrospun scaffolds can be processed post production for tissue engineering applications. Here we describe methods for spinning complex scaffolds (by consecutive spinning), for making thicker scaffolds (by multi-layering using heat or vapour annealing), for achieving sterility (aseptic production or sterilisation post production) and for achieving appropriate biomechanical properties.

조직 공학을위한 Electrospun의 Fibres의 Postproduction 처리

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

Tissue Engineering of a Human 3D in vitro Tumor Test System

Cancer is one of the leading causes of death worldwide. Current therapeutic strategies are predominantly developed in 2D culture systems, which inadequately reflect physiological conditions in vivo. Biological 3D matrices provide cells an environment in which cells can self-organize, allowing the study of tissue organization and cell differentiation. Such scaffolds can be seeded with a mixture of different cell types to study direct 3D cell-cell-interactions. To mimic the 3D complexity of cancer tumors, our group has developed a 3D in vitro tumor test system.	
	Our 3D tissue test system models the in vivo situation of malignant peripheral nerve sheath tumors (MPNSTs), which we established with our decellularized porcine jejunal segment derived biological vascularized scaffold (BioVaSc). In our model, we reseeded a modified BioVaSc matrix with primary fibroblasts, microvascular endothelial cells (mvECs) and the S462 tumor cell line. For static culture, the vascular structure of the BioVaSc is removed and the remaining scaffold is cut open on one side (Small Intestinal Submucosa SIS-Muc). The resulting matrix is then fixed between two metal rings (cell crowns).	
	Another option is to culture the cell-seeded SIS-Muc in a flow bioreactor system that exposes the cells to shear stress. Here, the bioreactor is connected to a peristaltic pump in a self-constructed incubator. A computer regulates the arterial oxygen and nutrient supply via parameters such as blood pressure, temperature, and flow rate. This setup allows for a dynamic culture with either pressure-regulated pulsatile or constant flow.	
	In this study, we could successfully establish both a static and dynamic 3D culture system for MPNSTs. The ability to model cancer tumors in a more natural 3D environment will enable the discovery, testing, and validation of future pharmaceuticals in a human-like model.

Tissue Engineering of a Human 3D in vitro Tumor Test System

Methods to create human 3D tumor tissues as test systems are described. These technologies are based on a decellularized Biological Vascularized Scaffold (BioVaSc), primary human cells and a tumor cell line, which can be cultured under static as well as under dynamic conditions in a flow bioreactor.

인간의 3 차원의 조직 공학<em&gt; 시험관</em&gt; 종양 테스트 시스템

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. Collagen has been extensively used for a wide range of biomedical applications and is considered a valid alternative to synthetic materials due to its inherent biocompatibility (i.e., low antigenicity, inflammation, and cytotoxic responses). However, the limited mechanical properties and the related low hand-ability of collagen gels have hampered their use as scaffold materials for vascular tissue engineering. Therefore, the rationale behind this work was first to engineer cellularized collagen gels into a tubular-shaped geometry and second to enhance smooth muscle cells driven reorganization of collagen matrix to obtain tissues stiff enough to be handled.
The strategy described here is based on the direct assembling of collagen and smooth muscle cells (construct) in a 3D cylindrical geometry with the use of a molding technique. This process requires a maturation period, during which the constructs are cultured in a bioreactor under static conditions (without applied external dynamic mechanical constraints) for 1 or 2 weeks. The &#8220;static bioreactor&#8221; provides a monitored and controlled sterile environment (pH, temperature, gas exchange, nutrient supply and waste removal) to the constructs. During culture period, thickness measurements were performed to evaluate the cells-driven remodeling of the collagen matrix, and glucose consumption and lactate production rates were measured to monitor the cells metabolic activity. Finally, mechanical and viscoelastic properties were assessed for the resulting tubular constructs. To this end, specific protocols and a focused know-how (manipulation, gripping, working in hydrated environment, and so on) were developed to characterize the engineered tissues.

In this work, we present a technique for the rapid fabrication of living vascular tissues by direct culturing of collagen, smooth muscle cells and endothelial cells. In addition, a new protocol for the mechanical characterization of engineered vascular tissues is described.

혈관 조직 재생을위한 엔지니어링 3D Cellularized 콜라겐 젤

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. ...

Preparation of Thermoresponsive Nanostructured Surfaces for Tissue Engineering

Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-immobilized surfaces for controlling cell adhesion and detachment were fabricated by the Langmuir-Schaefer method. Amphiphilic block copolymers composed of polystyrene and PIPAAm (St-IPAAms) were synthesized by reversible addition-fragmentation chain transfer (RAFT) radical polymerization. A chloroform solution of St-IPAAm molecules was gently dropped into a Langmuir-trough apparatus, and both barriers of the apparatus were moved horizontally to compress the film to regulate its density. Then, the St-IPAAm Langmuir film was horizontally transferred onto a hydrophobically modified glass substrate by a surface-fixed device. Atomic force microscopy images clearly revealed nanoscale sea-island structures on the surface. The strength, rate, and quality of cell adhesion and detachment on the prepared surface were modulated by changes in temperature across the lower critical solution temperature range of PIPAAm molecules. In addition, a two-dimensional cell structure (cell sheet) was successfully recovered on the optimized surfaces. These unique PIPAAm surfaces may be useful for controlling the strength of cell adhesion and detachment.

Nanoscaled sea-island surfaces composed of thermoresponsive block copolymers were fabricated by the Langmuir-Schaefer method for controlling spontaneous cell adhesion and detachment. Both the preparation of the surface and the adhesion and detachment of cells on the surface were visualized.

조직 공학 Thermoresponsive 나노 구조 표면의 제조

Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-immobilized surfaces for controlling cell adhesion and detachment were fabricated by the ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

에 내장 혈관 신생에 의하여 조직 공학<em&gt; 생체</em&gt; 조직 공학 상공 회의소

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

A Protocol for Decellularizing Mouse Cochleae for Inner Ear Tissue Engineering

In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current regenerative medicine strategies have focused on transplanting stem cells or genetic manipulation of surrounding support cells in the inner ear to encourage replacement of damaged stem cells to correct hearing loss. Yet, the extracellular matrix (ECM) may play a vital role in inducing and maintaining function of hair cells, and has not been well investigated. Using the cochlear ECM as a scaffold to grow adult stem cells may provide unique insights into how the composition and architecture of the extracellular environment aids cells in sustaining hearing function. Here we present a method for isolating and decellularizing cochleae from mice to use as scaffolds accepting perfused adult stem cells. In the current protocol, cochleae are isolated from euthanized mice, decellularized, and decalcified. Afterward, human Wharton&#39;s jelly cells (hWJCs) that were isolated from the umbilical cord were carefully perfused into each cochlea. The cochleae were used as bioreactors, and cells were cultured for 30 days before undergoing processing for analysis. Decellularized cochleae retained identifiable extracellular structures, but did not reveal the presence of cells or noticeable fragments of DNA. Cells perfused into the cochlea invaded most of the interior and exterior of the cochlea and grew without incident over a duration of 30 days. Thus, the current method can be used to study how cochlear ECM affects cell development and behavior.

The goal of this protocol is to demonstrate an effective method to decellularize and decalcify mouse cochleae for utilization as scaffolds for tissue engineering applications.

내가 조직 공학에 대 한 마우스 Cochleae Decellularizing에 대 한 프로토콜

In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current ...

Methods Article

심장 조직 공학을위한 모세관 력 리소그래피