저자는 시작 기금 및 서던 일리노이 대학, 의과 대학에서 외과 연구 보조금의 부서를 인정하고 싶습니다; 및 기념 의료 재단 보조금.

1. 수술 불러 참고 : IRB 승인이 ACL 재건 수술하는 동안 ACL 그루터기를 수집 얻었다. 사용되는 ACL 그루터기는 일반적으로 수술 폐기물로 폐기되었다으로 IRB 면제가 주어졌다. 20 명의 환자는 ACL 그루터기를 수집하는 데 사용되었다. <ol><li> 환자에게 기관의 프로토콜에 따라 전신 마취 및 수술 전 항생제를 관리 할 수​​ 있습니다. </li><li> 지혈대를 적용하고 수축기 혈압 이상 100 ㎎의 팽창. </li><li> 앙와위에서 환자를 놓다. 5mm의 포털에 대한 30 ~ 45 °의 굴곡에 무릎과 수평 외측 절개를 30 °에서 suprapatellar 주머니로 관절 경 트로 카메라를 삽입합니다. </li><li> 일상적인 진단 관절 경을 수행합니다. </li><li> 노치를 입력하고 찢어진 AC​​L에 막 활액막을 debride. </li><li> ACL 그루터기를 식별 할 수있는 4.5 mm 면도기 및 애블 레이터를 사용합니다. </li><li> 여러 개의 SPE를 수집하는 직선 쓴 짱구를 사용나머지 ACL 그루터기에서 cimens. </li><li> 멸균 용기에 생리 식염수에서 샘플을 유지한다. </li><li> 디코딩 및 실험실 시설에 최종 전달을 위해 병리에 견본을 보낸다. </li><li> 환자를 복구하고 기관의 프로토콜에 따라 수술 치료를 수행합니다. </li></ol> 2. 조직 소화 hACL 격리 <ol><li> 멸균 식염수 / PBS와 페트리 접시에 병리로부터 수신 ACL 인간 조직을 전송 (인산염 완충 염수). </li><li> 1~2밀리미터 3 조각으로 멸균 가위를 사용하여 조직을 말하다 및 식염수로 3 회 이상 다진 조직을 씻어. </li><li> DMEM/F-12 0.4 % 콜라게나 37 ℃에서 4 ~ 6 시​​간 동안 10 % 소 태아 혈청 (FBS), 1 % 페니실린 / 스트렙토 마이신과 보충 ((50 / 50), 1X) 매체와 다진 조직 다이제스트 </li><li> 5 분 동안 1,000 XG에서 세포를 원심 분리하고 배지에서 세포를 재현 탁. </li><li> 다음의 2 ~ 3 배의 배지로 세포를 씻어EED / 2 ~ 3 일 T-25 플라스크에 문화. </li></ol> 3. 세포 확장, 냉동 및 해동 <ol><li> T-25 플라스크에 2 ~ 3 일 배양 세포를 시각화하기 위해 광학 현미경을 사용합니다. </li><li> DMEM/F-12에서 37 ° C에서 세포를 10 % 소 태아 혈청 (FBS), 1 % 페니실린 / 스트렙토 마이신 (P / S)로 보충 ((50 / 50), 1X) 매체를 유지한다. 참고 : 세포 부착 및 표준 형태를 표시 할 때 미디어를 변경합니다. 세포를 관찰하는 광학 현미경을 사용합니다. </li><li> 7 일 (~ 1.3 X 10 6 세포 / 플라스크)에서 PBS에 합류 세포를 씻으십시오. 트립신 (0.5 g / L)을 추가하고 5 분 동안 37 ° C에서 세포를 품어. 트립신을 중화 일시 중단 세포에 미디어를 추가합니다. 세 개의 T-25 플라스크의 미디어 세포 혼합물을 나눈다. </li><li> 트립신 (0.5 g / L)를 추가, PBS로 합류 세포를 씻어 DMSO, FBS와 완전한 매체를 포함하는 매체를 동결에서 그들을 재현 탁의 (DMEM/F-12 중간 + 10 % FBS + 1 % P / S) 비 1시 2분 7초. </li><li> 저장세포가 더 긴 기간 동안 저장 될 경우 세포가 한 달 내에 사용하고, 액체 질소 될 경우 -80 ° C에서 저온 유리 병. </li><li> 재사용을 위해 37 ° C의 물을 욕조에 튜브를 해동. 참고 : 일반적으로 90 % -95 %의 세포가 남아있다. </li></ol> 4. 2-D 폴리 락트산 - 글리콜 산 (PLAGA) 비계 엔지니어링 조직의 제작 <ol><li> 20 ㎖ 섬광 유리 병에 12 ㎖의 디클로로 메탄에 PLAGA 1 g을 녹여 일정한 속도 (800 RPM)에서 8 시간 동안 솔루션을 소용돌이. 참고 :. 굽타 등 15 자세한 설명 </li><li> 불화 보호 종이 줄 지어 유리 페트리 접시에 용해 된 용액을 전송합니다. </li><li> 30 분 동안 진공 후드 아래에 페트리 접시를 놓습니다. 실온에서 하룻밤 후 -20 ° C에서 배양 접시를 남겨주세요. 고분자의 박막을 얻기 위해 용매의 완전한 증발을 보장하기 위해 동결 건조기의 배양 접시를 놓습니다. </li><li> 폴리머 Fi를 배치24 시간 동안 건조기에서 LMS. </li><li> 12mm 직경의 원형 디스크에 폴리머 필름을 잘라 필요할 때까지 데시 케이 터에 보관하십시오. </li></ol> 5. 주사 전자 현미경 (SEM) <ol><li> 세포 (50,000 세포 / 디스크)를 세척 파종 7 일 to 이후 PBS와 제어 TCPS 및 PLAGA 비계에 시드. </li><li> 세포를 고정 후 고정 용 0.1 M Cacodylate 버퍼에 2.5 % OSO 4 0.1 M Cacodylate 버퍼에 1.5 % 글루 타르 알데히드를 사용합니다. </li><li> 0.1 M Cacodylate 버퍼를 사용하여 고정 된 세포를 씻으십시오. 15 분마다, 밤새 헥사 메틸 디 실라 잔 (HMDS)의 추가 건조를 위해 직렬 탈수 에탄올 (50 %, 70 %, 80 %, 90 % 및 100 %)을 사용하여 고정 된 세포를 건조. </li><li> 버튼 밸브 SEM 스퍼터 코팅 시스템의 챔버를 배출하고 상판을 올린다. </li><li> 챔버에서 건조 된 샘플을 놓습니다. 상판을 낮 춥니 다. </li><li> 배기를 시작하는 펌프 컨트롤 노브를 전환합니다. </li><li> 오픈 아르곤 누출 밸브. 잠깐만 요진공 0.05 밀리바에 떨어 뜨리도록. </li><li> 금 / 팔라듐의 얇은 층 (0.13 NM)와 코트 건조 된 샘플. </li><li> 폐쇄 위치에 아르곤 누출 밸브를 돌려줍니다. </li><li> 오프에 컨트롤 노브를 전환합니다. </li><li> 버튼 밸브 챔버 환기와 상판을 올린다. </li><li> 샘플을 제거하고 24 시간 동안 건조기에 저장합니다. </li><li> 주사 전자 현미경으로 시료를 관찰한다. </li></ol> 6. 면역 형광 염색법 <ol><li> 세포 (50,000 세포 / 디스크)를 세척 파종 7 일 to 이후 PBS와 제어 TCPS 및 PLAGA 비계에 시드. </li><li> 10 분 동안 70 % 에탄올 (차가운)를 사용하여 세포를 고정한다. </li><li> PBS는 20 분 동안 0.05 % 트리톤 X-100을 갖는 1 % 소 혈청 알부민 (BSA)로 실온에서 고정 된 세포를 배양한다. </li><li> 20 분 동안 실온에서 1 % 트윈으로 샘플을 담금. </li><li> 단클론 마우스 안티-β-액틴 항체 (1:400)를 추가하고 overnigh 세포를 배양4 ° C에서 T </li><li> 0.05 % 트윈으로 세포를 씻으십시오. 세포를 이차 항체 (항 - 마우스 F (AB &#39;산양) 형광 프로브, 1:400으로 공액의 IgG 2 단편)을 추가하고, 실온에서 1 시간 동안 배양한다. </li><li> PBS로 세포를 씻으십시오. 핵 얼룩 세포를 얼룩과 글리세롤 80 %를 사용하여 마운트합니다. </li><li> 공 초점 현미경으로 세포를 관찰합니다. </li></ol>

<ol>
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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review+of+anterior+cruciate+ligament+reconstruction+with+autograft+compared+with+allograft.">A systematic review of anterior cruciate ligament reconstruction with autograft compared with allograft.</a> J Bone Joint Surg. 91, 2242-2450 (2009).</li><li>Murray, M. M., Martin, S. D., Martin, T. L., Spector, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+changes+in+the+human+anterior+cruciate+ligament+after+rupture.">Histological changes in the human anterior cruciate ligament after rupture.</a> J Bone Joint Surg Am. 82, 1387-1397 (2000).</li><li>Andrish, J., Holmes, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+synovial+fluid+on+fibroblasts+in+tissue+culture.">Effects of synovial fluid on fibroblasts in tissue culture.</a> Clin Orthop Relat Res. (138), 279-283 (1979).</li><li>Murray, M. M., Spindler, K. P., Ballard, P., 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=Enhanced+histologic+repair+in+a+central+wound+in+the+anterior+cruciate+ligamentwith+a+collagen-platelet-rich+plasma+scaffold.">Enhanced histologic repair in a central wound in the anterior cruciate ligamentwith a collagen-platelet-rich plasma scaffold.</a> J Orthop Res. 25, 1007-1017 (2007).</li><li>Carter, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anterior+cruciate+ligament+thermal+shrinkage.">Anterior cruciate ligament thermal shrinkage.</a> Clin Sports Med. 21, 693-700 (2002).</li><li>Perry, J. J., Higgins, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anterior+and+posterior+cruciate+ligament+rupture+after+thermal+treatment.">Anterior and posterior cruciate ligament rupture after thermal treatment.</a> Arthroscopy. 16, 732-736 (2000).</li><li>Cheng, M. T., Yang, H. W., Chen, T. H., Lee, O. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+multipotent+stem+cells+from+human+cruciate+ligaments.">Isolation and characterization of multipotent stem cells from human cruciate ligaments.</a> Cell Prolif. 42 (4), 448-460 (2009).</li><li>Matsumoto, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+human+anterior+cruciate+ligament-derived+vascular+stem+cells.">Isolation and characterization of human anterior cruciate ligament-derived vascular stem cells.</a> Stem Cells Dev. 21 (6), 859-872 (2012).</li></ol>

저자는 더 경쟁 재정적 이익이 없다는 것을 선언합니다.

이 hACL/2D 발판 연구의 주요 목적은 부분​​ ACL 눈물의 차 수리를 증가시키기 위해 패치에서 얻은 세포를 사용하는 것이 었습니다. 부분 ACL 눈물의 비수술 적 관리는 고정, 보조기, 진보적 인 재활 프로그램, 정기적 인 후속 평가 13,16,17의 짧은 기간을 포함 할 수있다. 그러나 많은 연구는 운동 선수의 보존 적 치료가 가난한 결과와 실패와 관련되어 있음을 보여줍니다. 버클리 등 알....

전방 십자 인대 (ACL)은 무릎 부상 일반적으로 관절 내 인대이다. 약 20 만 (ACL) 부상은 미국에서 매년보고됩니다. 정형 외과 재건 수술 1,2,3,4에 대한 ACL 부상 OPT가 발생 환자의 75 % 이상. 외과 적 수술은 종종 인해 본질적으로 가난한 치유의 가능성 (3)에 표시됩니다. 전방 십자 인대 재건술은 일반적으로자가 이식 또는 동종 이식 건을 이용하여 수행됩니다. 그들은 높은 성공률 및 차 봉합 수리, 다른 치료 옵션을 자랑으로자가 이식과 동종 이식은 재건을위한 황금 표준을 나타냅니다, 최대 94 % 5,6,7의 실패율을 보였다. ACL의 부분 눈물이 모든 ACL의 눈물 (8)의 28 %에 10 %를 나타냅니다. 전향 적 연구에서, 노이스 등. 더 ACL의 절반 이상에 영향을 미치는 부분 눈물을 가진 환자의 50 %가 후 ACL 부족을 완료하기 위해 비 진행 추정수술 적 치료 9. 다른 연구는 지속적인 불안정을보고하고 자신의 전 상해 활동 레벨 9,10,11,12,13로 돌아갈 수의 환자 미만 30 %와 기능을 감소. 치료 옵션이 제한됩니다 보수적 인 양식, 나머지 ACL의 열 수축, 또는 전방 십자 인대 재건술이 (가) 있습니다. 최근 증강 차 수리에 대한 관심이 증가하고있다. 이 기술은 차 봉합 수리 (14)을 향상시키기 위해 생물학적 제제를 사용합니다. 최근의 연구는 중간 엽 같은 hACL는, 이식 제한을 회피하기 위해 (31, 32)를 줄기 세포를 유도하지만, 이러한 세포의 타당성과 유효성은 아직 알 수없는 수확을 시도하고있다. 조직 공학 응용 프로그램에 대한 이상적인 셀 소스 hACL은 섬유 아세포의 세포를 유도 비 mesechymal 것으로 보인다. 현재 연구는 적절한 매트릭스 재료와 설계 비계에 대한 세포 소스를 식별에 초점을 맞추고 있습니다. 그것은 스와로 폐기 될 찢어진 AC​​L에 대한 표준 절차재건 수술 중 rgical 폐기물은, 그러나,이 손상된 인대는 ACL 교체를 설계에 이상적인 조직을 개발하고 강화하는 데 필요한 세포의 획득을위한 품질의 소스가 될 수 있습니다. 우리 연구소는 이러한 수확 hACL 유래 세포의 체외 확장을위한 프로토콜을 개발했습니다. hACL 유래 세포 주입 설계 2D 매트릭스를 사용하여, 우리는 잠재적으로 부분 ACL 수리를 확장하고 찢어진 인대를 강화 할 수있는 패치를 설계했다.

<table border="0" cellpadding="0" cellspacing="0" style="width:1004px;" width="1003">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:139px;">Company</td>
			<td style="width:143px;">Catalog Number</td>
			<td style="width:507px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Petri-dish</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">08-757-103C</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:216px;">Phosphate Buffered Saline (PBS)</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">BP3994</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Collagenase</td>
			<td style="width:139px;">Gibco</td>
			<td style="width:143px;">17018-029</td>
			<td>Store at 40C</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">DMEM/F-12</td>
			<td style="width:139px;">Cellgro</td>
			<td style="width:143px;">10-092-CV</td>
			<td>Store at 40C. Warm in 370C water bath before use.</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Fetal Bovine Serum (FBS)</td>
			<td style="width:139px;">Gibco</td>
			<td style="width:143px;">10082</td>
			<td>Store at -800C</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Penicillin/Streptomycin</td>
			<td style="width:139px;">Lonza</td>
			<td style="width:143px;">17-602E</td>
			<td>Store at 40C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Centrifuge Tubes- 15 ml</td>
			<td style="width:139px;">Corning</td>
			<td style="width:143px;">430790</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">T-25 flasks</td>
			<td style="width:139px;">BD Falcon</td>
			<td style="width:143px;">3013</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Trypsin-Versene mixture</td>
			<td style="width:139px;">Lonza</td>
			<td style="width:143px;">17-161E</td>
			<td>Store at 40C. Warm in 370C water bath before use.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DMSO</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">BP231-100</td>
			<td>Combustible liquid. Can cause skin, eye and respiratory tract irritation.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cryogenic vials</td>
			<td style="width:139px;">Corning</td>
			<td style="width:143px;">430489</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">PLAGA</td>
			<td style="width:139px;">Purac Biomaterials</td>
			<td style="width:143px;">Purasorb PLG8523</td>
			<td>Store at -800C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">TCPS disks</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">12-545-82</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:216px;">Dichloromethane</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">AC36423-0010</td>
			<td>Possible cancer hazard. Store in a dry, cool place.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Bytac paper</td>
			<td style="width:139px;">Saint gobin performance plastics</td>
			<td style="width:143px;">1420652</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scintillation vial</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td>03-339-21G</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Bovine Serum Albumin (BSA)</td>
			<td style="width:139px;">Sigma Aldrich</td>
			<td style="width:143px;">A7906</td>
			<td>Store at 40C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tween</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">BP337-500</td>
			<td></td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:216px;">mouse Anti-&#946;-actin antibody (10 Ab)</td>
			<td style="width:139px;">Sigma Aldrich</td>
			<td style="width:143px;">A5441</td>
			<td>Store at -200C</td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:216px;">goat-anti-mouse antibody (20 Ab)</td>
			<td style="width:139px;">Cell Signaling</td>
			<td style="width:143px;">4408</td>
			<td>Store at -200C</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Hoechst dye</td>
			<td style="width:139px;">Sigma Aldrich</td>
			<td style="width:143px;">14530</td>
			<td>Store at -200C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glycerol</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">BP229-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glutaraldehyde</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">BP2547-1</td>
			<td>Toxic by inhalation and if swallowed. Causes burns by all exposure routes.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hexamethyldisilazane</td>
			<td style="width:139px;">Fisher Scientific</td>
			<td style="width:143px;">AC43085-1000</td>
			<td>Flammable liquid and vapor. Causes burns by all exposure routes.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sterile Scissors</td>
			<td style="width:139px;">McKesson</td>
			<td style="width:143px;">25-716</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Centrifuge</td>
			<td style="width:139px;">Eppendorf</td>
			<td style="width:143px;">5804R</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Light Microscope</td>
			<td style="width:139px;">Olympus</td>
			<td style="width:143px;">CK40</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Water Bath</td>
			<td style="width:139px;">Thermo scientific</td>
			<td style="width:143px;">2845</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vortex</td>
			<td style="width:139px;">Labnet&#160;</td>
			<td style="width:143px;">VX-200</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glass Petri plates</td>
			<td style="width:139px;">fisher Scientific</td>
			<td style="width:143px;">S31473</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Acu-Punch</td>
			<td style="width:139px;">Acuderm Inc.</td>
			<td style="width:143px;">P1225</td>
			<td>Acu-Punch was used to cut 12mm disks</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cacodylate buffer</td>
			<td style="width:139px;">Sigma Aldrich</td>
			<td style="width:143px;">97068</td>
			<td>Flammable liquid, carcinogen and irritant.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Osmium tetraoxide</td>
			<td style="width:139px;">Sigma Aldrich</td>
			<td style="width:143px;">201030</td>
			<td>Highly toxic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Triton X-100</td>
			<td style="width:139px;">Sigma Aldrich</td>
			<td style="width:143px;">T8787</td>
			<td>Harmful if swallowed</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ethanol</td>
			<td style="width:139px;">Decon Labs</td>
			<td style="width:143px;">2705</td>
			<td>Keep away from heat, sparks, flame and other form of ignition.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">General/regional Anesthesia</td>
			<td style="width:139px;">Amphastar pharmaceuticals</td>
			<td style="width:143px;"></td>
			<td>1% Lidocaine, 0.25% Bupivacaine,Anesthetic agents for induction and maintenance</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Antibiotics</td>
			<td style="width:139px;">Hospira</td>
			<td>0409-0805-01</td>
			<td>Ancef 1g i.v.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Arthroscopy trocar</td>
			<td style="width:139px;">Smith and Nephew</td>
			<td style="width:143px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Arthroscopy Camera</td>
			<td style="width:139px;">Smith and Nephew</td>
			<td style="width:143px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Arthroscopic grasper and bitter</td>
			<td style="width:139px;">Arthrex&#160;</td>
			<td style="width:143px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">4.5mm shaver</td>
			<td style="width:139px;">Arthrex&#160;</td>
			<td style="width:143px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Interference Screws</td>
			<td style="width:139px;">Arthrex&#160;</td>
			<td style="width:143px;"></td>
			<td>stainless steel screws</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sputter Coater</td>
			<td style="width:139px;">Polaron</td>
			<td style="width:143px;">E5400</td>
			<td></td>
		</tr>
	</tbody>
</table>

surgical retrieval, isolation and in vitro expansion of human anterior cruciate ligament-derived cells for tissue engineering applications

ACL에 부상을 활성 개인에 일반적으로 발생하는 문제입니다. 기능과 안정성에 악영향을 생 역학적 결함이 관절 내 무릎 인대 리드 부분도 눈물. 열 변형, 단일 번들 수리, 완전한 재건, 네이티브 인대의 손상 부분의 재건 : 부분 ACL 눈물의 치료를 위해 현재 옵션은 보존 적, 보수적 인 경영에서 다음과 같은 여러 수술 옵션에 이르기까지 다양합니다. 연구는 거의 모든 경우, 관리는 지속적으로 우수한 것으로에 대한 하나의 방법을 증명하고있다, 많은 경우에 환자는 지속적인 불안정과 다른 동반 질환을 설명하기 위해 계속합니다. 본 연구의 목표는 부분적으로 찢어 ACL의 수선에서 구현 될 수있는 조직 공학적 패치의 개발에 활용 가능성 셀 소스를 식별하는 것이다. 새로운 프로토콜은 patie에서 파생 된 세포의 확장을 위해 개발 된전방 십자 인대 재건술을받은 국세청. 세포를 분리하기 위해 ACL 재구성 동안 획득 다진 hACL 조직은 콜라게나 제 용액에서 분해시켰다. 확장은 10 % 태아 소 혈청 (FBS) 및 1 % 페니실린 / 스트렙토 마이신 (P / S)이 보충 된 DMEM/F12 배지를 사용하여 실시 하였다. 세포는 그 다음 DMSO, FBS와 팽창 매체로 이루어진 동결 배지에서 -80 º C의 또는 액체 질소에 보관 하였다. 해동 후, hACL 유래 세포는 조직 공학적 지지체, PLAGA (폴리 락트산 - 코 - 글리콜 산) 및 제어 조직 배양 폴리스티렌 (TCPS)에 접종 하였다. 7 일 후에, SEM은 제어 TCPS 대 PLAGA 셀룰러 밀착성과 비교하기 위해 수행 하였다. 세포 형태는 면역 형광 염색법을 사용하여 평가 하였다. SEM (주사 전자 현미경) 현미경은 세포가 성장하고 PLAGA 및 TCPS 양면에 부착하고 7 일째하여 전체 표면에 컨 플루 언트 된 것을 보여 주었다. 면역 형광 염색이 아닌 일반 강조 형태 학적을 보여 주었다양면에 알 패턴. 이 기술은 ACL 재생 및 재건에 응용 프로그램에 대한 약속한다.

외과 검색, 조직의 소화와 인간의 전방 십자 인대 (hACL) 유래 세포의 분리를위한 작업 모델은 그림 1에 나타내었다. 외식에서 마이그레이션 및 T-25 플라스크에 부착 된 세포를. 이 세포는 3 일 동안 배양 한 후 광학 현미경 (그림 2)에서 가시화되었다. 합류 단층은 하루 7을 얻었다. 건강, 생존 세포의 존재가 hACL 유래 세포를 성공적으로 검색 및 문화를 지적했다. <p class="jo...

Watch this Scientific Journal Video about Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications at JoVE.com

Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications

전방 십자 인대 (ACL)의 부분적인 눈물을 복구하는 패치 향후 애플리케이션의 경우, 인간의 ACL 유래 세포는 체외에서 확장 및 조직 공학적 지지체에 성장 재건 과정 중에 얻은 조직에서 분리되었다. 세포 접착 및 형태학이어서 지지체 표면에 생체 적합성을 확인하기 위하여 실시 하였다.

외과 검색, 분리 및<em&gt; 체외</em&gt; 조직 공학 응용 프로그램에 대한 인간의 전방 십자 인대에서 파생 된 세포의 확장

Injury to the ACL is a commonly encountered problem in active individuals. Even partial tears of this intra-articular knee ligament lead to biomechanical ...

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JoVE Journal

Bioengineering

Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications

11.4K 조회수.  Southern Illinois University School of Medicine.  전방 십자 인대 (ACL)의 부분적인 눈물을 복구하는 패치 향후 애플리케이션의 경우, 인간의 ACL 유래 세포는 체외에서 확장 및 조직 공학적 지지체에 성장 재건 과정 중에 얻은 조직에서 분리되었다. 세포 접착 및 형태학이어서 지지체 표면에 생체 적합성을 확인하기 위하여 실시 하였다. 

비디오: Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications

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 ...

연구

생체공학

Athymic Rat Model for Evaluation of Engineered Anterior Cruciate Ligament Grafts

Anterior cruciate ligament (ACL) rupture is a common ligamentous injury that often requires surgery because the ACL does not heal well without intervention. Current treatment strategies include ligament reconstruction with either autograft or allograft, which each have their associated limitations. Thus, there is interest in designing a tissue-engineered graft for use in ACL reconstruction. We describe the fabrication of an electrospun polymer graft for use in ACL tissue engineering. This polycaprolactone graft is biocompatible, biodegradable, porous, and is comprised of aligned fibers. Because an animal model is necessary to evaluate such a graft, this paper describes an intra-articular athymic rat model of ACL reconstruction that can be used to evaluate engineered grafts, including those seeded with xenogeneic cells. Representative histology and biomechanical testing results at 16 weeks postoperatively are presented, with grafts tested immediately post-implantation and contralateral native ACLs serving as controls. The present study provides a reproducible animal model with which to evaluate tissue engineered ACL grafts, and demonstrates the potential of a regenerative medicine approach to treatment of ACL rupture.

Animal models are important tools for the evaluation of tissue-engineered grafts. This paper presents the protocol for preparing an electrospun biodegradable polymer graft for use in anterior cruciate ligament tissue engineering, as well as a surgical protocol for implantation in a rat model.

엔지니어링 전방 십자 인대 이식의 평가 흉선 쥐 모델

Anterior cruciate ligament (ACL) rupture is a common ligamentous injury that often requires surgery because the ACL does not heal well without ...

Eye Irritation Test (EIT) for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial (RhCE) Tissue Model

To comply with the Seventh Amendment to the EU Cosmetics Directive and EU REACH legislation, validated non-animal alternative methods for reliable and accurate assessment of ocular toxicity in man are needed. To address this need, we have developed an eye irritation test (EIT) which utilizes a three dimensional reconstructed human cornea-like epithelial (RhCE) tissue model that is based on normal human cells. The EIT is able to separate ocular&#160;irritants and corrosives (GHS Categories 1 and 2 combined) and those that do not require labeling (GHS No Category). The test utilizes two separate protocols, one designed for liquid chemicals and a second, similar protocol for solid test articles. The EIT prediction model uses a single exposure period (30 min for liquids, 6 hr for solids) and a single tissue viability cut-off (60.0% as determined by the MTT assay). Based on the results for 83 chemicals (44 liquids and 39 solids) EIT achieved 95.5/68.2/ and 81.8% sensitivity/specificity and accuracy (SS&#38;A) for liquids, 100.0/68.4/ and 84.6% SS&#38;A for solids, and 97.6/68.3/ and 83.1% for overall SS&#38;A. The EIT will contribute significantly to classifying the ocular irritation potential of a wide range of liquid and solid chemicals without the use of animals to meet regulatory testing requirements. The EpiOcular EIT method was implemented in 2015 into the OECD Test Guidelines as TG 492.

Eye Irritation Test EIT for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial RhCE Tissue Model

We have developed an eye irritation test which utilizes a three dimensional reconstructed human cornea-like epithelial (RhCE) tissue model. The test is able to discriminate between ocular irritant and corrosive materials (GHS Categories 1 and 2 combined) and those that do not require labeling (GHS No Category).

복원 된 인간의 각막 모양의 상피 (RHCE) 조직 모델을 사용하여 눈에 자극성 화학 물질의 위험 식별을위한 눈 자극 시험 (EIT)

To comply with the Seventh Amendment to the EU Cosmetics Directive and EU REACH legislation, validated non-animal alternative methods for reliable and ...

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from traumatic injury, disease and various congenital, genetic and acquired conditions are quite common. Tissue engineering and regenerative medicine technologies have enormous potential to provide a therapeutic solution. However, utilization of biologically relevant animal models in combination with longitudinal assessments of pertinent functional measures are critical to the development of improved regenerative therapeutics for treatment of VML-like injuries. In that regard, a commercial muscle lever system can be used to measure length, tension, force and velocity parameters in skeletal muscle. We used this system, in conjunction with a high power, bi-phase stimulator, to measure in vivo force production in response to activation of the anterior crural compartment of the rat hindlimb. We have previously used this equipment to assess the functional impact of VML injury on the tibialis anterior (TA) muscle, as well as the extent of functional recovery following treatment of the injured TA muscle with our tissue engineered muscle repair (TEMR) technology. For such studies, the left foot of an anaesthetized rat is securely anchored to a footplate linked to a servomotor, and the common peroneal nerve is stimulated by two percutaneous needle electrodes to elicit muscle contraction and dorsiflexion of the foot. The peroneal nerve stimulation-induced muscle contraction is measured over a range of stimulation frequencies (1-200 Hz), to ensure an eventual plateau in force production that allows for an accurate determination of peak tetanic force. In addition to evaluation of the extent of VML injury as well as the degree of functional recovery following treatment, this methodology can be easily applied to study diverse aspects of muscle physiology and pathophysiology. Such an approach should assist with the more rational development of improved therapeutics for muscle repair and regeneration.

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

We describe an in vivo protocol to measure dorsiflexion of the foot following stimulation of the peroneal nerve and contraction of the anterior crural compartment of the rat hindlimb. Such measurements are an indispensable translational tool for evaluating skeletal muscle pathology and tissue engineering approaches to muscle repair and regeneration.

의 응용 프로그램<em&gt; 생체</em&gt; 조직 평가를위한 쥐 앞정 전방의 기능 테스트는 골격근 수리 엔지니어링

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from ...

Melt Electrospinning Writing of Three-dimensional Poly(&#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology with great potential for biomedical applications. The technique facilitates the direct deposition of biocompatible polymer fibers to fabricate well-ordered scaffolds in the sub-micron to micro scale range. The establishment of a stable, viscoelastic, polymer jet between a spinneret and a collector is achieved using an applied voltage and can be direct-written. A significant benefit of a typical porous scaffold is a high surface-to-volume ratio which provides increased effective adhesion sites for cell attachment and growth. Controlling the printing process by fine-tuning the system parameters enables high reproducibility in the quality of the printed scaffolds. It also provides a flexible manufacturing platform for users to tailor the morphological structures of the scaffolds to their specific requirements. For this purpose, we present a protocol to obtain different fiber diameters using melt electrospinning writing (MEW) with a guided amendment of the parameters, including flow rate, voltage and collection speed. Furthermore, we demonstrate how to optimize the jet, discuss often experienced technical challenges, explain troubleshooting techniques and showcase a wide range of printable scaffold architectures.

Melt Electrospinning Writing of Three-dimensional Poly(&amp;#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This protocol serves as a comprehensive guideline to fabricate scaffolds via electrospinning with polymer melts in a direct writing mode. We systematically outline the process and define the appropriate parameter settings for achieving targeted scaffold architectures.

조직 공학 응용 프로그램에 대 한 제어 형태학과 3 차원 Poly(ε-caprolactone) 장비의 전기 글 녹아

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology ...

Isolation of Mesenchymal Stem Cells from Human Alveolar Periosteum and Effects of Vitamin D on Osteogenic Activity of Periosteum-derived Cells

Mesenchymal stem cells (MSCs) are present in a variety of tissues and can be differentiated into numerous cell types, including osteoblasts. Among the dental sources of MSCs, the periosteum is an easily accessible tissue, which has been identified to contain MSCs in the cambium layer. However, this source has not yet been widely studied.
Vitamin D3 and 1,25-(OH)2D3 have been demonstrated to stimulate in vitro differentiation of MSCs into osteoblasts. In addition, vitamin C facilitates collagen formation and bone cell growth. However, no study has yet investigated the effects of Vitamin D3 and Vitamin C on MSCs.
Here, we present a method of isolating MSCs from human alveolar periosteum and examine the hypothesis that 1,25-(OH)2D3 may exert an osteoinductive effect on these cells. We also investigate the presence of MSCs in the human alveolar periosteum and assess stem cell adhesion and proliferation. To assess the ability of vitamin C (as a control) and various concentrations of 1,25-(OH)2D3 (10&#8722;10, 10&#8722;9, 10&#8722;8, and 10&#8722;7 M) to alter key mRNA biomarkers in isolated MSCs mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), core binding factor alpha-1 (CBFA1), collagen-1, and osteocalcin (OCN) are measured using real-time polymerase chain reaction (RT-PCR).

We present a protocol to investigate the mRNA expression biomarkers of periosteum-derived cells (PDCs) induced by vitamin C (vitamin C) and 1,25-dihydroxy vitamin D [1,25-(OH)2D3]. In addition, we evaluate the ability of PDCs to differentiate into osteocytes, chondrocytes, and adipocytes.

인간의 치경과 파생 셀 Osteogenic 활동에 비타민 D의 효과에서 중간 엽 줄기 세포의 분리

Mesenchymal stem cells (MSCs) are present in a variety of tissues and can be differentiated into numerous cell types, including osteoblasts. Among the ...

Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications

The autologous, synthetic, and animal-derived grafts currently used as scaffolds for tissue replacement have limitations due to low availability, poor biocompatibility, and cost. Plant tissues have favorable characteristics that make them uniquely suited for use as scaffolds, such as high surface area, excellent water transport and retention, interconnected porosity, preexisting vascular networks, and a wide range of mechanical properties. Two successful methods of plant decellularization for tissue engineering applications are described here. The first method is based on detergent baths to remove cellular matter, which is similar to previously established methods used to clear mammalian tissues. The second is a detergent-free method adapted from a protocol that isolates leaf vasculature and involves the use of a heated bleach and salt bath to clear the leaves and stems. Both methods yield scaffolds with comparable mechanical properties and low cellular metabolic impact, thus allowing the user to select the protocol which better suits their intended application.

Here we present, and contrast two protocols used to decellularize plant tissues: a detergent-based approach and a detergent-free approach. Both methods leave behind the extracellular matrix of the plant tissues used, which can then be utilized as scaffolds for tissue engineering applications.

Decellularization 식물 조직의 조직 공학 응용 프로그램에 대 한 두 가지 방법

The autologous, synthetic, and animal-derived grafts currently used as scaffolds for tissue replacement have limitations due to low availability, poor ...

Production of Autologous Platelet-Rich Plasma for Boosting In Vitro Human Fibroblast Expansion

There is currently great clinical interest in the use of autologous fibroblasts for skin repair. In most cases, culture of skin cells in vitro is required. However, cell culture using xenogenic or allogenic culture media has some disadvantages (i.e., risk of infectious agent transmission or slow cell expansion). Here, an autologous culture system is developed for the expansion of human skin fibroblast cells in vitro using a patient&#8217;s own platelet-rich plasma (PRP). Human dermal fibroblasts are isolated from the patient while undergoing abdominoplasty. Cultures are followed for up to 7 days using a medium supplemented with either fetal bovine serum (FBS) or PRP. Blood cell content in PRP preparations, proliferation, and fibroblast differentiation are assessed. This protocol describes the method for obtaining a standardized, non-activated preparation of PRP using a dedicated medical device. The preparation requires only a medical device (CuteCell-PRP) and centrifuge. This device is suitable under sufficient medical practice conditions and is a one-step, apyrogenic, and sterile closed system that requires a single, soft spin centrifugation of 1,500 x g for 5 min. After centrifugation, the blood components are separated, and the platelet-rich plasma is easily collected. This device allows a quick, consistent, and standardized preparation of PRP that can be used as a cell culture supplement for in vitro expansion of human cells. The PRP obtained here contains a 1.5-fold platelet concentration compared to whole blood together, with a preferential removal of red and white blood cells. It is shown that PRP presents a boosting effect in cell proliferation compared to FBS (7.7x) and that fibroblasts are activated upon PRP treatment.

This protocol presents a device that produces PRP to boost the in vitro expansion of cells in a 100% autologous fibroblast culture system.

In Vitro 인간 섬유아세포 확장을 촉진하기 위한 자가 혈소판 풍부 혈장 생산

There is currently great clinical interest in the use of autologous fibroblasts for skin repair. In most cases, culture of skin cells in vitro is ...

Anterior Segment Organ Culture Platform for Tracking Open Globe Injuries and Therapeutic Performance

Open globe injuries have poor visual outcomes, often resulting in permanent loss of vision. This is partly due to an extended delay between injury and medical intervention in rural environments and military medicine applications where ophthalmic care is not readily available. Untreated injuries are susceptible to infection after the eye has lost its watertight seal, as well as loss of tissue viability due to intraocular hypotension. Therapeutics to temporarily seal open globe injuries, if properly developed, may be able to restore intraocular pressure and prevent infection until proper ophthalmic care is possible. To facilitate product development, detailed here is the use of an anterior segment organ culture open globe injury platform for tracking therapeutic performance for at least 72 h post-injury. Porcine anterior segment tissue can be maintained in custom-designed organ culture dishes and held at physiological intraocular pressure. Puncture injuries can be created with a pneumatic-powered system capable of generating injury sizes up to 4.5 mm in diameter, similar to military-relevant injury sizes. Loss of intraocular pressure can be observed for 72 h post-injury confirming proper injury induction and loss of the eye&#39;s watertight seal. Therapeutic performance can be tracked by application to the eye after injury induction and then tracking intraocular pressure for multiple days. Further, the anterior segment injury model is applicable to widely used methods for functionally and biologically tracking anterior segment physiology, such as assessing transparency, ocular mechanics, corneal epithelium health, and tissue viability. Overall, the method described here is a necessary next step toward developing biomaterial therapeutics for temporarily sealing open globe injuries when ophthalmic care is not readily available.

Open globe eye injuries may go untreated for multiple days in rural or military-relevant scenarios, resulting in blindness. Therapeutics are needed to minimize loss of vision. Here, we detail an organ culture open globe injury model. With this model, potential therapeutics for stabilizing these injuries can be properly evaluated.

오픈 글로브 부상 및 치료 성능을 추적하기위한 전방 세그먼트 장기 문화 플랫폼

Open globe injuries have poor visual outcomes, often resulting in permanent loss of vision. This is partly due to an extended delay between injury and ...

Fibroblast Derived Human Engineered Connective Tissue for Screening Applications

Fibroblasts are phenotypically highly dynamic cells, which quickly transdifferentiate into myofibroblasts in response to biochemical and biomechanical stimuli. The current understanding of fibrotic processes, including cardiac fibrosis, remains poor, which hampers the development of new anti-fibrotic therapies. Controllable and reliable human model systems are crucial for a better understanding of fibrosis pathology. This is a highly reproducible and scalable protocol to generate engineered connective tissues (ECT) in a 48-well casting plate to facilitate studies of fibroblasts and the pathophysiology of fibrotic tissue in a 3-dimensional (3D) environment. ECT are generated around the poles with tunable stiffness, allowing for studies under a defined biomechanical load. Under the defined loading conditions, phenotypic adaptations controlled by cell-matrix interactions can be studied. Parallel testing is feasible in the 48-well format with the opportunity for the time-course analysis of multiple parameters, such as tissue compaction and contraction against the load. From these parameters, biomechanical properties such as tissue stiffness and elasticity can be studied.

Presented here is a protocol to generate engineered connective tissues for a parallel culture of 48 tissues in a multi-well plate with double poles, suitable for mechanistic studies, disease modeling, and screening applications. The protocol is compatible with fibroblasts from different organs and species and is exemplified here with human primary cardiac fibroblasts.

섬유아세포, 선별 애플리케이션을 위한 인간 공학형 결합 조직

Fibroblasts are phenotypically highly dynamic cells, which quickly transdifferentiate into myofibroblasts in response to biochemical and biomechanical ...