Name: processing of human cardiac tissue toward extracellular matrix self-assembling hydrogel for in vitro and in vivo applications
Uploaded: 2017-12-04T14:00:00
Description: Watch this Scientific Journal Video about Processing of Human Cardiac Tissue Toward Extracellular Matrix Self-assembling Hydrogel for In Vitro and In Vivo Applications at JoVE.com

Undersøgelse-protokollen i overensstemmelse med de etiske principper, der er skitseret i Helsinki-erklæringen. Patienter informeret samtykke til brug af væv i forskningsøjemed, og processen med væv samling blev godkendt af institutionelle Review Board og etiske komité i Charité - Universitätsmedizin Berlin (EA4/028/12).

Undersøgelse-protokollen er i overensstemmelse med de etiske principper, der er skitseret i Helsinki-erklæringen og blev godkendt af den institutionelle review board og etik af Charité medicinske universitet. Alle patienter der skrevet, informeret samtykke til brug af hjerte væv i eksperimentelle undersøgelser.
1. forberedelse af menneskelige myokardiet for skæring
<ol><li>Få venstre ventrikulære myokardiet (størrelse varierer afhængigt af typen kirurgi) direkte fra operationsstuen...

<ol>
	<li>Elliott, R., Hoehn, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+Commercial+Porcine+Skin+for+Wound+Dressings.">Use of Commercial Porcine Skin for Wound Dressings.</a> Plastic and reconstructive surgery. 52 (4), 401-405 (1973).</li><li>Rienks, M., Papageorgiou, A. -. P., Frangogiannis, N. G., Heymans, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myocardial+Extracellular+Matrix:+An+Ever-Changing+and+Diverse+Entity.">Myocardial Extracellular Matrix: An Ever-Changing and Diverse Entity.</a> Circulation Research. 114 (5), 872-888 (2014).</li><li>Prabhu, S. D., Frangogiannis, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Biological+Basis+for+Cardiac+Repair+After+Myocardial+Infarction.">The Biological Basis for Cardiac Repair After Myocardial Infarction.</a> Circulation Research. 119 (1), 91-112 (2016).</li><li>Boopathy, A. V., Martinez, M. D., Smith, A. W., Brown, M. E., Garcia, A. J., Davis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intramyocardial+Delivery+of+Notch+Ligand-Containing+Hydrogels+Improves+Cardiac+Function+and+Angiogenesis+Following+Infarction.">Intramyocardial Delivery of Notch Ligand-Containing Hydrogels Improves Cardiac Function and Angiogenesis Following Infarction.</a> Tissue Eng Part A. 21 (17-18), 2315-2322 (2015).</li><li>Gaetani, R., Yin, C., 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=Cardiac+derived+extracellular+matrix+enhances+cardiogenic+properties+of+human+cardiac+progenitor+cells.">Cardiac derived extracellular matrix enhances cardiogenic properties of human cardiac progenitor cells.</a> Cell Transplant. , (2015).</li><li>Kraehenbuehl, T. P., Ferreira, L. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+embryonic+stem+cell-derived+microvascular+grafts+for+cardiac+tissue+preservation+after+myocardial+infarction.">Human embryonic stem cell-derived microvascular grafts for cardiac tissue preservation after myocardial infarction.</a> Biomaterials. 32 (4), 1102-1109 (2011).</li><li>Zhang, J., Klos, M., 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=Extracellular+matrix+promotes+highly+efficient+cardiac+differentiation+of+human+pluripotent+stem+cells:+The+matrix+sandwich+method.">Extracellular matrix promotes highly efficient cardiac differentiation of human pluripotent stem cells: The matrix sandwich method.</a> Circulation Research. 111 (9), 1125-1136 (2012).</li><li>Fong, A. H., Romero-L&#243;pez, M., 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=Three-Dimensional+Adult+Cardiac+Extracellular+Matrix+Promotes+Maturation+of+Human+Induced+Pluripotent+Stem+Cell-Derived+Cardiomyocytes.">Three-Dimensional Adult Cardiac Extracellular Matrix Promotes Maturation of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.</a> Tissue Engineering Part A. 22 (15-16), 1016-1025 (2016).</li><li>DeQuach, J. A., Mezzano, 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=Simple+and+High+Yielding+Method+for+Preparing+Tissue+Specific+Extracellular+Matrix+Coatings+for+Cell+Culture.">Simple and High Yielding Method for Preparing Tissue Specific Extracellular Matrix Coatings for Cell Culture.</a> PLoS ONE. 5 (9), e13039 (2010).</li><li>Saldin, L. T., Cramer, M. C., Velankar, S. S., White, L. J., Badylak, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+matrix+hydrogels+from+decellularized+tissues:+Structure+and+function.">Extracellular matrix hydrogels from decellularized tissues: Structure and function.</a> Acta Biomaterialia. 49, 1-15 (2017).</li><li>Tukmachev, D., Forostyak, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injectable+extracellular+matrix+hydrogels+as+scaffolds+for+spinal+cord+injury+repair.">Injectable extracellular matrix hydrogels as scaffolds for spinal cord injury repair.</a> Tissue Eng Part A. , (2016).</li><li>Freytes, D. O., Martin, J., Velankar, S. S., Lee, A. S., Badylak, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+and+rheological+characterization+of+a+gel+form+of+the+porcine+urinary+bladder+matrix.">Preparation and rheological characterization of a gel form of the porcine urinary bladder matrix.</a> Biomaterials. 29 (11), 1630-1637 (2008).</li><li>Singelyn, J. M., Sundaramurthy, 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=Catheter-deliverable+hydrogel+derived+from+decellularized+ventricular+extracellular+matrix+increases+endogenous+cardiomyocytes+and+preserves+cardiac+function+post-myocardial+infarction.">Catheter-deliverable hydrogel derived from decellularized ventricular extracellular matrix increases endogenous cardiomyocytes and preserves cardiac function post-myocardial infarction.</a> Journal of the American College of Cardiology. 59 (8), 751-763 (2012).</li><li>Wainwright, J. M., Czajka, C. 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=Preparation+of+cardiac+extracellular+matrix+from+an+intact+porcine+heart.">Preparation of cardiac extracellular matrix from an intact porcine heart.</a> Tissue Eng Part C Methods. 16 (3), 525-532 (2010).</li><li>Ott, H. C., Matthiesen, T. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perfusion-decellularized+matrix:+using+nature's+platform+to+engineer+a+bioartificial+heart.">Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart.</a> Nature Medicine. 14, 213-221 (2008).</li><li>Oberwallner, B., Anic, B. 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=Human+cardiac+extracellular+matrix+supports+myocardial+lineage+commitment+of+pluripotent+stem+cells.">Human cardiac extracellular matrix supports myocardial lineage commitment of pluripotent stem cells.</a> Eur J Cardiothorac Surg. 47, 416-425 (2015).</li><li>Oberwallner, B., Brodarac, 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=Preparation+of+cardiac+extracellular+matrix+scaffolds+by+decellularization+of+human+myocardium.">Preparation of cardiac extracellular matrix scaffolds by decellularization of human myocardium.</a> Journal of Biomedical Materials Research Part A. 102 (9), 3263-3272 (2014).</li><li>Kappler, B., Anic, 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=The+cytoprotective+capacity+of+processed+human+cardiac+extracellular+matrix.">The cytoprotective capacity of processed human cardiac extracellular matrix.</a> Journal of Materials Science: Materials in Medicine. , (2016).</li><li>Bashey, R. I., Martinez-Hernandez, A., Jimenez, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation,+characterization,+and+localization+of+cardiac+collagen+type+VI.+Associations+with+other+extracellular+matrix+components.">Isolation, characterization, and localization of cardiac collagen type VI. Associations with other extracellular matrix components.</a> Circulation Research. 70 (5), (1992).</li><li>Wu, J., Ravikumar, P., Nguyen, K. T., Hsia, C. C. W., Hong, Y., Gorler, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+protection+by+inhalation+of+exogenous+solubilized+extracellular+matrix.">Lung protection by inhalation of exogenous solubilized extracellular matrix.</a> PLOS ONE. 12 (2), e0171165 (2017).</li><li>Chen, W. C. W., Wang, Z., 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=Decellularized+zebrafish+cardiac+extracellular+matrix+induces+mammalian+heart+regeneration.">Decellularized zebrafish cardiac extracellular matrix induces mammalian heart regeneration.</a> Science Advances. 2 (11), e1600844 (2016).</li><li>Godier-Furn&#233;mont, A. F. G., Martens, T. 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=Composite+scaffold+provides+a+cell+delivery+platform+for+cardiovascular+repair.">Composite scaffold provides a cell delivery platform for cardiovascular repair.</a> Proceedings of the National Academy of Sciences of the United States of America. 108 (19), 7974-7979 (2011).</li><li>Sarig, U., Sarig, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Natural+myocardial+ECM+patch+drives+cardiac+progenitor+based+restoration+even+after+scarring.">Natural myocardial ECM patch drives cardiac progenitor based restoration even after scarring.</a> Acta Biomaterialia. 44, 209-220 (2016).</li><li>Singelyn, J. M., DeQuach, J. A., Seif-Naraghi, S. B., Littlefield, R. B., Schup-Magoffin, P. J., Christman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Naturally+derived+myocardial+matrix+as+an+injectable+scaffold+for+cardiac+tissue+engineering.">Naturally derived myocardial matrix as an injectable scaffold for cardiac tissue engineering.</a> Biomaterials. 30 (29), 5409-5416 (2009).</li></ol>

Når du forbereder menneskelige Myokardie ECM, målet er at opnå følgende: fjernelse af relevante immunogen cellular materiale, bevarelse af ECM integritet og bioactivity, sterilitet, ikke-giftighed af slutproduktet, GMP-processen kompatibilitet, og egnethed af produktet til en given anvendelse med hensyn til håndtering. Ved at kombinere vores 3-trins decellularization protokol med yderligere forarbejdning til mikropartikler eller selvsamlende hydrogel, menneskelige hjerte ECM materiale er fremstillet som besidder sæ...

Den ekstracellulære matrix (ECM) giver ikke kun strukturfondene støtter men er også vigtig for biologiske celle og væv funktion1. I hjertet deltager ECM i reguleringen af patofysiologiske svar som fibrose, betændelse, angiogenese, cardiomyocyte kontraktile funktion og levedygtighed og bosiddende stamfader celle skæbne. Ud over sin primære komponenter - fiber glykoproteiner, glycosaminoglycans og proteoglycaner - indeholder den et væld af udskilles vækstfaktorer, cytokiner og hindeagtige vesikler indeholdende nukleinsyrer og proteiner2,3.
Det er for nylig blevet klart, at acellulær ECM præparater ikke er kun uvurderlig for at studere celle-matrix interaktioner, men også for potentielle terapeutiske celle-baserede programmer. Betydningen af at skabe et passende miljø til terapeutiske celle produkter eller manipuleret væv er nu bredt anerkendt. Man har forsøgt at kombinere celle suspensioner eller aktive forbindelser med definerede biopolymeric hydrogels4,5,6 eller protein cocktails udskilles af murine sarkom celler (dvs., Matrigel, Geltrex) 7. men førstnævnte har begrænset bioactivity, sidstnævnte er problematisk i GMP-grade processer, og begge mangler væv-specifikke bioactivity i hjertets ECM (cECM)8,9,10, 11,12,13.
Decellularization af myokardiet er tidligere blevet udført af perfusion af hele hjertet via koronar Vaskulaturen14,15. Mens dette er muligt i dyrenes hjerter, er intakt menneskers hjerter sjældent tilgængelige. Derfor, en nedsænkning proces, der giver mulighed for håndtering af vævsprøver i operationsstuen var begunstiget. Vores "3-trins" protokol indeholder 3 separate inkubation skridt nemlig lysis, oploesning og DNA fjernelse. Det giver menneskers Myokardie ECM med stort set bevarede protein og glykosaminoglykan sammensætning16,17. Disse cECM skiver mulighed for in vitro- undersøgelser af celle-matrix interaktioner men er dårligt egnet til potentielle menneskelige skala terapeutiske anvendelsesmuligheder. Fremstillingsprocessen var derefter udvidet til at producere enten frysetørret cECM mikropartikler eller en cECM hydrogel18.
Denne protokol giver mulighed for decellularization af menneskelige myokardiet fremstillet af kirurgisk prøver, bevare de væsentligste komponenter i Myokardie ekstracellulære matrix (ECM) og deres biologiske aktivitet. Denne protokol kan anbefales når menneskelige hjerte ECM med bevaret væv-specifikke bioactivity er nødvendig for eksperimentelle undersøgelser af celle-matrix interaktioner, eller når et passende miljø er nødvendige for cellebaserede Myokardie regenerering tilgange. I princippet er det også muligt at tilpasse denne protokol til GMP-grade betingelser, således at brugen af forarbejdede cECM skal være muligt i fremtiden terapeutiske anvendelsesmuligheder.

<table>
	<tbody>
		<tr>
			<td>Balance</td>
			<td>DR Precisa, Dietikon, Switzerland</td>
			<td>Precisa XR 205SM</td>
			<td></td>
		</tr>
		<tr>
			<td>Blades Nr.10 Skalpell Nr.3</td>
			<td>InstrumenteNRW, Erftstadt, Germany</td>
			<td>SK-10-004</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell culture plates (6-well)</td>
			<td>Greiner, Frickenhausen, Germany</td>
			<td>657160</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryostat CM</td>
			<td>Leica, Wetzlar, Germany</td>
			<td>3050S</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Carl Roth, Karlsruhe, Germany</td>
			<td>8043.3</td>
			<td></td>
		</tr>
		<tr>
			<td>Eppendorf reaction tubes (1.5 or 2 ml)</td>
			<td>Greiner, Frickenhausen, Germany</td>
			<td>616201, 623201</td>
			<td></td>
		</tr>
		<tr>
			<td>Falcon 15ml, 50ml</td>
			<td>Greiner, Frickenhausen, Germany</td>
			<td>188271, 227270</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Biochrome, Berlin, Germany</td>
			<td>S 0115</td>
			<td></td>
		</tr>
		<tr>
			<td>Freeze Dry System</td>
			<td>Labconco, Kansas City, USA</td>
			<td>7670520</td>
			<td></td>
		</tr>
		<tr>
			<td>Freezer (-80&#176;C)</td>
			<td>Thermo Scientific, Waltham, MA, USA</td>
			<td>Forma 900 Series</td>
			<td></td>
		</tr>
		<tr>
			<td>HCl</td>
			<td>Carl Roth, Karlsruhe, Germany</td>
			<td>281.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Microtome Blades Type 819</td>
			<td>Leica, Wetzlar, Germany</td>
			<td>14035838925</td>
			<td></td>
		</tr>
		<tr>
			<td>Minilys Homogeniser</td>
			<td>PEQLAB Biotechnologie GmbH, Erlangen, Germany</td>
			<td>91-PCSM</td>
			<td></td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td>Carl Roth, Karlsruhe, Germany</td>
			<td>K021.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Nystatin</td>
			<td>PAN Biotech, Aidenbach, Germany</td>
			<td>P06-07800</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Thermo Scientific, Waltham, MA, USA</td>
			<td>14190-094</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin/streptomycin</td>
			<td>Life Technologies, Darmstadt, Germany</td>
			<td>15140122</td>
			<td></td>
		</tr>
		<tr>
			<td>Pepsin</td>
			<td>Sigma-Aldrich, Taufkirchen, Germany</td>
			<td>P6887-1G</td>
			<td></td>
		</tr>
		<tr>
			<td>Precellys Keramik-Kit 1.4 mm</td>
			<td>Peqlab Biotechnolgie, Erlangen, Germany</td>
			<td>91-PCS-CK14</td>
			<td></td>
		</tr>
		<tr>
			<td>Rotamax 120 Plate shaker</td>
			<td>Heidolph, Schwabach, Germany</td>
			<td>544-41200-00</td>
			<td></td>
		</tr>
		<tr>
			<td>SDS</td>
			<td>Carl Roth, Karlsruhe, Germany</td>
			<td>CN30.3</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>Leica, Wetzlar, Germany</td>
			<td>M125</td>
			<td></td>
		</tr>
		<tr>
			<td>Steriflip-GP, 0,22 &#181;m</td>
			<td>Merck Millipore, Darmstadt, Germany</td>
			<td>SCGP00525</td>
			<td></td>
		</tr>
		<tr>
			<td>Stuart analogue rocker &#38; roller mixers</td>
			<td>Sigma-Aldrich, Taufkirchen, Germany</td>
			<td>Z675113-1EA</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue Tek O.C.T compound</td>
			<td>Hartenstein, Wurzburg, Germany</td>
			<td>TTEK</td>
			<td></td>
		</tr>
		<tr>
			<td>Transfusion set 200&#181;m</td>
			<td>Sarstedt, N&#252;mbrecht, Germany</td>
			<td>798.200.500</td>
			<td></td>
		</tr>
		<tr>
			<td>TRIS</td>
			<td>Carl Roth, Karlsruhe, Germany</td>
			<td>5429.3</td>
			<td></td>
		</tr>
		<tr>
			<td>vedena Skalpellgriff Fig. 3, Standard, 125 mm</td>
			<td>Medical Highlights, Rohrdorf, Germany</td>
			<td>CV102-003</td>
			<td></td>
		</tr>
		<tr>
			<td>Vortex-Genie2</td>
			<td>Scientific Industry, New York, USA</td>
			<td>SI-0256</td>
			<td></td>
		</tr>
	</tbody>
</table>

processing of human cardiac tissue toward extracellular matrix self-assembling hydrogel for in vitro and in vivo applications

Acellulær ekstracellulære matrix præparater er nyttige til at studere celle-matrix interaktioner og lette regenerativ celle terapi programmer. Flere kommercielle ekstracellulære matrix produkter fås som hydrogels eller membraner, men disse er ikke i besidelse væv-specifikke biologiske aktivitet. Fordi perfusion decellularization ikke er normalt muligt med menneskelige hjerte væv, udviklede vi en 3-trins fordybelse decellularization proces. Menneskelige Myokardie skiver indkøbt under operationen behandles først med vaskemiddel-fri hyperosmolær lysisbuffer, efterfulgt af inkubation med Ioniske vaskemiddel, sodium dodecyl sulfat, og processen er færdig ved at udnytte den iboende DNase aktivitet af føtal bovint serum. Denne teknik resulterer i celle-gratis plader af cardiac ekstracellulære matrix med stort set bevaret fibrøst væv arkitektur og polymer sammensætning, som blev vist til at give specifikke miljømæssige stikord til hjerte cellepopulationer og pluripotente stamceller celler. Hjerte ekstracellulære matrix ark kan derefter yderligere forarbejdes til et microparticle pulver uden yderligere kemiske ændringer, eller, via kortsigtede pepsin fordøjelse, ind i en selvsamlende hjerte ekstracellulære matrix hydrogel med bevaret bioactivity.

3-trins protokol til decellularization af menneskelige myokardiet præsenteres her resultaterne i næsten fuldstændig fjernelse af cellulære materiale, samtidig bevare ECM nøglekomponenter og den fibrillar struktur af ECM. Efter decellularization fremgår grov fjernelse af celler fra vævet af ændringen i farve (figur 1A). Histologiske analyse med H & E og Masson Trichrome pletter afslørede den fuldstændige mangel på resterende intakt celler (<strong c...

Watch this Scientific Journal Video about Processing of Human Cardiac Tissue Toward Extracellular Matrix Self-assembling Hydrogel for In Vitro and In Vivo Applications at JoVE.com

Processing of Human Cardiac Tissue Toward Extracellular Matrix Self-assembling Hydrogel for In Vitro and In Vivo Applications

Denne protokol beskriver den komplette decellularization af menneskelige myokardiet samtidig bevare sin ekstracellulære matrix komponenter. Videreforarbejdning af ekstracellulære matrix resulterer i produktionen af mikropartikler og en cytoprotective selvsamlende hydrogel.

Behandling af menneskelige hjerte væv mod ekstracellulære Matrix selv samle Hydrogel for In Vitro og In Vivo applikationer

Acellular extracellular matrix preparations are useful for studying cell-matrix interactions and facilitate regenerative cell therapy applications. ...

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Developmental Biology

Processing of Human Cardiac Tissue Toward Extracellular Matrix Self-assembling Hydrogel for In Vitro and In Vivo Applications

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Characterization of Thymic Settling Progenitors in the Mouse Embryo Using In Vivo and In Vitro Assays

Karakterisering af tymisk Bundfældning Progenitorer i Mouse Embryo Brug<em&gt; In vivo</em&gt; Og<em&gt; In vitro</em&gt; Analyser

Characterizing thymic settling progenitors is important to understand the pre-thymic stages of T cell development, essential to devise strategies for T ...

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

Analyse af zebrafisk Larver Skeletmuskel Integritet med Evans blå farvestof

The zebrafish model is an emerging system for the study of neuromuscular disorders.  In the study of neuromuscular diseases, the integrity of the muscle ...

Isolation of Perivascular Multipotent Precursor Cell Populations from Human Cardiac Tissue

Isolering af Perivaskulær multipotente precursorcelle populationer fra humant hjertevæv

Multipotent mesenchymal stem/stromal cells (MSC) were conventionally isolated, through their plastic adherence, from primary tissue digests whilst their ...

Directed Differentiation of Primitive and Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells

Instrueret differentiering af Primitive og endelige hæmatopoietisk ophav fra humane pluripotente stamceller

One of the major goals for regenerative medicine is the generation and maintenance of hematopoietic stem cells (HSCs) derived from human pluripotent stem ...

Live Imaging of Mouse Secondary Palate Fusion

The fusion of the secondary palatal shelves to form the intact secondary palate is a key process in mammalian development and its disruption can lead to ...

Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids

Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids

Immuno-fluorescerende mærkning af mikrotubuli og Centrosomal proteiner i Ex Vivo tarmens væv og 3D In Vitro tarm Organoids

The advent of 3D in vitro organoids that mimic the in vivo tissue architecture and morphogenesis has greatly advanced the ability to study key biological ...

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

Celle sammenlægning Assays til at evaluere bindingen af Drosophila Notch med Trans-ligander og dens hæmning af Cis-ligander

Notch signaling is an evolutionarily conserved cell-cell communication system used broadly in animal development and adult maintenance. Interaction of the ...

Isolation of Human Endometrial Stromal Cells for In Vitro Decidualization

Isolation of Human Endometrial Stromal Cells for In Vitro Decidualization

Isolering af menneskelige endometrie stromale celler til In vitro- Decidualization

The differentiation of human endometrial stromal cells (HESC) from fibroblast-like appearance into secretory decidua is a transformation required for ...

Isolation of Rat Adipose Tissue Mesenchymal Stem Cells for Differentiation into Insulin-producing Cells

Isolering af rottefedtvæv Mesenkymale stamceller til differentiering i insulinproducerende celler

Mesenchymal stem cells (MSCs)-especially those isolated from adipose tissue (Ad-MSCs)-have gained special attention as a renewable, abundant source of ...

Development of Organoids from Mouse Pituitary as In Vitro Model to Explore Pituitary Stem Cell Biology

Development of Organoids from Mouse Pituitary as In Vitro Model to Explore Pituitary Stem Cell Biology

Udvikling af organoider fra musegravyse som in vitro-model til at udforske hypofysestamcellebiologi

The pituitary is the master endocrine gland regulating key physiological processes, including body growth, metabolism, sexual maturation, reproduction, ...