Name: stencil micropatterning of human pluripotent stem cells for probing spatial organization of differentiation fates
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Description: Watch this Scientific Journal Video about Stencil Micropatterning of Human Pluripotent Stem Cells for Probing Spatial Organization of Differentiation Fates at JoVE.com

Start up Zuschuss (R-397-000-192-133) und ETPL Gap Fund (R-397-000-198-592) Diese Arbeit wird von NUS unterstützt. GS ist ein NUS Forschungswissenschaftler. Autoren möchte Dr. Jiangwa Xing für ihre technische Unterstützung auf Zelle Mikro danken.

Hinweis: Dieses Protokoll beschreibt die Herstellung von PDMS Schablone mit 1000 um Muster durch Laserschneiden und Mikrostrukturierung der hESC Linie, H9 die PDMS-Schablone verwenden. 1. Konstruktion und Herstellung von PDMS-Schablone zur Mikro <ol><li> Gestalten Sie das Schablonieren Blatt mit Durchgangslöchern der gewünschten Geometrie und Größe (zB 1000 um Kreise) und die Schablone Dichtung mit Computer-Aided - Design - Software - 10. </li><li> Laser-schneiden S...

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Die Herstellung von Mikro Schablonen Stencil Mikrostrukturierung ist eine ideale Methode HPSC Mikro zur Untersuchung Nische vermittelte Differenzierung Strukturierung zu erzeugen. Der entscheidende Vorteil der Schablonenmuster über andere Mikrostrukturierungstechniken, wie Mikrokontaktdrucken und Photostrukturierungs, ist, dass sie nicht die Oberflächenmodifikation erforderlich ist und kann auf herkömmliche TCPS Substraten implementiert werden. Daher optimiert Ku...

Humanen pluripotenten Stammzellen (hPSCs), einschließlich embryonaler Stammzellen (hES-Zellen) und induzierten pluripotenten Stammzellen (hiPSCs) werden in der regenerativen Medizin sowie experimentelle Modellierung von normalen und erkrankten Organogenese aufgrund ihrer Differenzierungspotential in Zelllinien aller weit ausgebeutet drei Keimschichten 1,2. Die Differenzierung Schicksale von hPSCs sind sehr empfindlich auf lokale Umweltfaktoren , die 1 autokrine oder parakrine modulieren können Signalisierung sowie Mechanotransduktion Prozesse durch körperliche Signale vermittelt 3-5. Zelle Mikro umfasst eine Reihe von Techniken , die räumlich zu organisieren , um die Geometrie und die Position einer Zellpopulation als Mittel entwickelt wurden 3 die lokale zelluläre Mikroumgebung zu steuern, wie Zell-Zell - Wechselwirkungen 6 und Zell-Matrix - Wechselwirkungen. Im Rahmen der hPSCs wurde Zelle Mikro wichtige Erkenntnisse darüber zu gewinnen, wie beschäftigt Nischen abhängenent autokrine Signalisieren moduliert hESC Pluripotenz-Differenzierung Entscheidungen 7 und Organisation in der frühen embryonalen Differenzierungsmuster 6. 2D- und 3D mikro hPSCs wurden verwendet , um die Koloniegröße von mehrzelligen Mustern zu steuern, die wiederum beeinflusst Differenzierungs Entscheidungen in die drei Keimschichten 8,9. Wir haben vielzelligen HPSC Mikromuster verwendet , um das Ausmaß der Zell-Zell- und Zell-Matrix - Interaktionen innerhalb einer HPSC Kolonie zu modulieren , zu untersuchen , wie Integrin-E-Cadherin - Übersprechens 10 Anlass zu Zellschicksal Heterogenität geben kann. Die Demonstrationen aus den oben genannten Berichten eröffnen neue Wege in Richtung der Anwendung von mehrzelligen Mikro von hPSCs als experimentelle Modelle für Medikamententoxizität Screening auf Entwicklungsstörungen 11, die Wirkung von Wachstumsfaktoren und Hormone während der Gewebe oder Organentwicklung zu studieren, und die Bildung von zu entwirren Gewebemuster. Eine Vielzahl von Zell micropatterning - Techniken wurden von Falconnet et als prüft entwickelt. al. 12 , aber nur eine Handvoll, wie Mikrokontaktdrucken 7,8,13, Mikrowell - Kultur 14,15, Photostrukturierungs 6 und microstencils 16 wurden mit hPSCs erfolgreich umgesetzt. Die Herausforderung mit Mikro hPSCs liegt in ihrer Verletzlichkeit und einer stringenten Bedingung von spezifischen extrazellulären Matrices (ECM) und die Wachstumsbedingungen für die Zellhaftung und Überleben. Für 2D HPSC Muster ist Mikrokontaktdrucken eine der gebräuchlichsten Methoden HPSC Mikromuster auf Gewebekultur und Glassubstrate 13 zu erzeugen. Das Verfahren kann gemeinsam Muster ECM in HPSC Kultur verwendet werden, einschließlich Laminin und Basalmembran-Matrices wie Matrigel. Allerdings erfordert es in der Regel einen zweistufigen Beschichtungsverfahren unterstützt von Poly-D-Lysine und benötigt spezifische inerten atmosphärischen und Feuchtigkeitsbedingungen stabil ECM Mikromuster zu machen für hPSCs auf 6,13 zu befestigen. Die wichtigste Überlegung jedes Mikro Methode ist, ob die Oberflächenmodifikation Regime HPSC-Klebstoff ECM-Muster in der gewünschten geometrischen Auflösung bei gleichzeitiger Minimierung unspezifischer Zellbindung an die Umgebung erzeugen kann. Hier berichten wir die Verwendung von Schablonenmikro als einfaches Verfahren HPSC Mikro ohne zusätzliche Oberflächenmodifikationsschritte vor der Erzeugung von Klebstoff ECM Muster zu erzeugen, um hPSCs zu befestigen auf. Die Zell Schablone besteht aus einer dünnen Membran, beispielsweise Polydimethylsiloxan (PDMS) -Folie, mit micron Durchgangslöchern bis Millimetergröße auf ein Substrat Zellkultur abzudichtenden physikalisch ECM Beschichtungen und anschließend beimpft hPSCs enthalten. Als Schablonenmuster durch physikalisches Zurückhalten der Position arbeitet, wo HPSC kann an dem darunterliegenden ECM beschichtete Substrat direkt zuzugreifen und diese befestigen, ist dieses Verfahren kompatibel mit verschiedenen Substraten, die HPSC Kulturen unterstützen kann. Die einzigen requirement ist, dass die Wahl des Schablonenmaterial eine reversible Dichtung mit dem Substrat bilden kann. Diese Substrate umfassen herkömmliche Gewebekultur - Polystyrol (TCPS) 17, Liganden konjugiert Substrate 18 sowie elastomere Substrate mit abstimmbaren Steifigkeit (z. B. PDMS) 19. Dieses Verfahren ermöglicht auch das Beschichten von verschiedenen ECM, wie Vitronectin (oder VTN - Protein), Laminin und Basalmembran Matrices (zB Matrigel und Geltrax) für die richtige Befestigung und die Differenzierung von hPSCs zu ermöglichen. Deshalb können wir für eine bestimmte HPSC Linie optimierte ECM-Substrat-Konfigurationen Transfer zum Schablone für eine optimale Zell-Matrix-Adhäsion, das Überleben und die Differenzierung Mikrostrukturierung. Kürzlich wurde ein ähnliches Verfahren auch hepatische Differenzierung von Mikro hESCs unter Verwendung von Poly (methylmethacrylat) (PMMA) Mikroschablonenanordnungen 16 zu leiten berichtet. Zell Schablonen können aus verschiedenen Materialien hergestellt werden, einschließlich erfülltals 20,21, Poly (p-xylylen) Polymere 22,23, PMMA und 16 am häufigsten, PDMS 24-28. Silicon und Poly (p-xylylen) Polymere Schablonen erfordern direkte Ätzen der Durchgangslöcher mit Spezialgeräten 20-23, was deren Zugänglichkeit für die biologische Anwender einschränkt. PDMS Schablonen können durch verschiedene Verfahren hergestellt werden , in Abhängigkeit von der Merkmalsgröße benötigt, die typischerweise von 3 &amp; mgr; m bis 2.000 &amp; mgr; m reicht 11,26-29. Wenn kleine Features gewünscht werden, dünne stenciling Blätter können durch Pressformen PDMS pre-Polymer auf einem mikrostrukturierten Silizium Vorlage mit Reliefs der Mikromuster 28 hergestellt werden. Für Funktionen&gt; 1000 &amp; mgr; m, bietet ein CO 2 Laser - Cutter eine einfache und kostengünstige Methode , um direkt die Muster auf einem bereits gegossenen PDMS Blatt während Schablonenherstellung geschnitten. Die Rezyklierbarkeit von PDMS Schablonen macht sie auch kosteneffektiv eine Reihe von Experimenten mit ausreichender Konsistenz durchzuführen. <p class = &quot;jove_content&quot;&gt; Hier präsentieren wir die detaillierte Methodik für die Herstellung eines PDMS Schablone mit 1000 um Funktionen durch Laserschneiden und die Erzeugung von hESC Mikro. Diese hESC Mikro wurden verwendet , um das Ausmaß der Integrin und E-Cadherin vermittelte Verwachsungen innerhalb einer zusammenhängenden hESC Kolonie zu modulieren , um zu untersuchen , wie räumliche Polarisierung der Zelladhäsion führte Zellschicksal Heterogenität 10.

<table border="0" cellpadding="0" cellspacing="0" width="948">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">&#160;2 mm thick PDMS sheet</td>
			<td style="width:191px;">Specialty Silicone Products Inc., USA</td>
			<td style="width:119px;">SSPM823-.005</td>
			<td style="width:423px;">Used to form reservoir for stencil</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">120-150 &#956;m thick PDMS sheet</td>
			<td style="width:191px;">Specialty Silicone Products Inc., USA</td>
			<td style="width:119px;">SSPM823-.040</td>
			<td style="width:423px;">Used to form stencil&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">60 mm petri dish</td>
			<td style="width:191px;">&#160;Nunc Nunclon Delta</td>
			<td style="width:119px;">150326</td>
			<td style="width:423px;">Substrate for micropatterning</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Accutase</td>
			<td style="width:191px;">Accutase, Merck Millipore, Singapore</td>
			<td style="width:119px;">SCR005</td>
			<td style="width:423px;">Enzyme to break H9 Cells into single cells</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Activin&#160;&#160;</td>
			<td style="width:191px;">R&#38;D Systems, Singapore</td>
			<td style="width:119px;">338-AC-010</td>
			<td style="width:423px;">Growth factor for H9 differentiation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">BMP4&#160;</td>
			<td style="width:191px;">R&#38;D Systems, Singapore</td>
			<td style="width:119px;">338-BP-010</td>
			<td style="width:423px;">Growth factor for H9 differentiation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Plasma system&#160;</td>
			<td style="width:191px;">&#160;Femto Science, Korea</td>
			<td style="width:119px;">CUTE-MP</td>
			<td style="width:423px;">For plasma oxidation of stencil</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dispase</td>
			<td style="width:191px;">StemCell&#8482; Technologies, Singapore</td>
			<td style="width:119px;">7923</td>
			<td style="width:423px;">Enzyme used to weaken the cell-ECM adhesion during passaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DMEM/F12</td>
			<td style="width:191px;">GIBCO, USA</td>
			<td style="width:119px;">11330032</td>
			<td style="width:423px;">Basal medium for H9 cells</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">FGF2</td>
			<td style="width:191px;">&#160;R&#38;D Systems, Singapore</td>
			<td style="width:119px;">233&#8211;FB&#8211;025</td>
			<td style="width:423px;">Growth factor for H9 differentiation</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">H9 Cell line</td>
			<td style="width:191px;">WiCell Research Institute, Inc., USA</td>
			<td style="width:119px;">WA09</td>
			<td style="width:423px;">Human embryonic stem cells</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">hESC-qualified basement membrane matrix</td>
			<td style="width:191px;">Matrigel, BD Biosciences, Singapore</td>
			<td style="width:119px;">354277</td>
			<td style="width:423px;">Extra-cellular matrix coating to support growth of H9 cells</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Inverted microscope</td>
			<td style="width:191px;">Leica Microsystems, Singapore</td>
			<td style="width:119px;">DMi1</td>
			<td style="width:423px;">For capturing bright-field images</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Laser cutter</td>
			<td style="width:191px;">Epilog Helix 24 Laser System</td>
			<td style="width:119px;"></td>
			<td style="width:423px;">Used to generate through holes in PDMS sheet</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">mTeSR&#8482;1 medium&#160;</td>
			<td style="width:191px;">StemCell&#8482; Technologies, Singapore</td>
			<td style="width:119px;">5850</td>
			<td style="width:423px;">Maintainence medium for H9 cells</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">PDMS&#160;</td>
			<td style="width:191px;">SYLGARD&#174; 184, Dow Corning Co., USA</td>
			<td style="width:119px;">3097358-1004</td>
			<td style="width:423px;">Used for sticking the PDMS stencil and reservior</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">ROCKi Y27632</td>
			<td style="width:191px;">Calbiochem, Merck Millipore, Singapore</td>
			<td style="width:119px;">688000</td>
			<td style="width:423px;">Maintains H9 cells as single cells&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">STEMdiff&#8482; APEL&#8482; medium&#160;</td>
			<td style="width:191px;">StemCell&#8482; Technologies, Singapore</td>
			<td style="width:119px;">5210</td>
			<td style="width:423px;">Differentiation medium for H9 cells</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Polyethylene terephthalate film</td>
			<td style="width:191px;">SureMark Singapore</td>
			<td style="width:119px;">SQ-6633</td>
			<td style="width:423px;">Used to form stencil&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Cell culture compatible non-ionic surfactant</td>
			<td style="width:191px;">Pluronic acid F-127, Sigma, Singapore</td>
			<td style="width:119px;">P2443</td>
			<td style="width:423px;">Passivating reagent to repel cell adhesion in non-micropatterned substrates</td>
		</tr>
	</tbody>
</table>

stencil micropatterning of human pluripotent stem cells for probing spatial organization of differentiation fates

Human pluripotent stem cells (hPSCs), including embryonic stem cells and induced pluripotent stem cells, have the intrinsic ability to differentiate into all three germ layers. This makes them an attractive cell source for regenerative medicine and experimental modeling of normal and diseased organogenesis. However, the differentiation of hPSCs in vitro is heterogeneous and spatially disordered. Cell micropatterning technologies potentially offer the means to spatially control stem cell microenvironments and organize the resultant differentiation fates. Micropatterning hPSCs needs to take into account the stringent requirements for hPSC survival and maintenance. Here, we describe stencil micropatterning as a method that is highly compatible with hPSCs. hPSC micropatterns are specified by the geometries of the cell stencil through-holes, which physically confine the locations where hPSCs can access and attach to the underlying extracellular matrix-coated substrate. Due to this mode of operation, there is greater flexibility to use substrates that can adequately support hPSCs as compared to other cell micropatterning methods. We also highlight critical steps for the successful generation of hPSC micropatterns. As an example, we demonstrate that stencil micropatterning of hPSCs can be used to modulate spatial polarization of cell-cell and cell-matrix adhesions, which in turn determines mesoendoderm differentiation patterns. This simple and robust method to micropattern hPSCs widens the prospects of establishing experimental models to investigate tissue organization and patterning during early embryonic development.

In diesem Papier beschreiben wir die Herstellung einer Zelle Schablone durch einen Laserschneider 1000 um Merkmale zu erzeugen. Die Schablone wurde aus 2 Teilen: einem dünnen Blech Schablonieren (ungefähr 100-200 &amp; mgr; m dick), enthaltend die Mikrodurchgangslöchern und einem PDMS Dichtung die ECM Beschichtungslösung oder Zellsuspension zu enthalten. Hier, 127 &amp; mgr; m und 2 mm dicken handelsüblichen PDMS Bleche wurden als Schablonieren Folie und Dichtung verwendet wurden. A...

Watch this Scientific Journal Video about Stencil Micropatterning of Human Pluripotent Stem Cells for Probing Spatial Organization of Differentiation Fates at JoVE.com

Stencil Micropatterning of Human Pluripotent Stem Cells for Probing Spatial Organization of Differentiation Fates

Humanen pluripotenten Stammzellen (hPSCs) haben die intrinsische Fähigkeit zu differenzieren und sich selbst zu organisieren in verschiedene Gewebemuster; obwohl dies erfordert die Darstellung der räumlichen Umweltgradienten. Wir präsentieren Schablone Mikro als einfache und robuste Methode biochemische und mechanische Gradienten zu erzeugen, für HPSC Differenzierungsmuster zu steuern.

Stencil Mikrostrukturierung von humanen pluripotenten Stammzellen für die Sondierung räumliche Organisation Differenzierung Fates

Human pluripotent stem cells (hPSCs), including embryonic stem cells and induced pluripotent stem cells, have the intrinsic ability to differentiate into ...

The overall goal of this stencil-based micropatterning method, for human pluripotent stem cells, is to provide a simple and robust means to generate spatial environmental gradients that control stem cell differentiation patterns. This method can help you answer key questions in tissue patterning using a human stem cell model, such as how spatial gradients in cell-cell and cell matrix adhesion forces can help to organize stem cell differentiation fates. The advantage of this method is that it does not require additional surface modification to generate cell adhesive ECM patterns for cells to attach to.This means the method is compatible with various ECM and substrate configurations, specific for each cell line. Visual demonstration of this method is critical, as the micropatterning steps are difficult to learn. Because it requires precise and careful handling of the stencil and the cells.For this procedure, the PTMS stencils must already be designed and fabricated. Likewise, have human EFC cells growing on basement membrane matrix, such that the colonies will be ready for seeding later in the procedure. To begin, load a dish with 700 microliters of 70%analytical-grade ethanol, in ultra-pure water.Then, place the PDMS stencil on the solution, and set the dish into a bio-safety cabinet to dry out overnight. The next day, check that no bubbles have formed under the stencil. Then, field the dish until the cells can be seeded onto the stencil.Prepare an aliquot of human ESC-quality basement membrane matrix by adding 16.7 milliliters of DMF12, to make a 1.5x solution and keep it on ice. Treat the substrate with 100 watts of oxygen plasma for 90 seconds. Don't open the dish until it is inside the plasma chamber, and quickly cover it up after the treatment.This treatment will prevent bubbles from forming when the substrate is immersed in solution. Now, cover the stencil with 450 microliters of prepared 1.5x basement membrane matrix solution. Then, seal the dish with parafilm and incubate it for five hours.Before proceeding, check over the six well plates of prepared HESC colonies. Identify and aspirate off colonies of differentiated cells that have lost their typical HESC morphology. They should all appear rounded, have a tightly-packed epithelial morphology, and have a high nucleus-to-cytoplasm ratio with prominent nucleoli.Then, wash each well of plated cells with two milliliters of DMF12 two times. After the second wash, add one milliliter of digestive enzymes to each well, such as accutase and incubate the plate for eight minutes at 37 degrees Celsius. After the incubation, tap the plate gently to detach all the colonies from the substrate.Next, rinse each well with at least four milliliters of media for every milliliter of digestive enzyme applied to the well, and collect the cell suspensions into a single 15 milliliter conical tube. Centrifuge the cells at 200 g for three minutes at room temperature to form a cell pellet. Then aspirate the supernatant, and re-suspend the cells in 400 microliters of HESC maintenance media, supplemented with 10 micromolar of rock inhibitor.Triturate the cells gently three times to break up the clumps. Next, correct the suspensions'density. Mix it well, and make a 200 microliter 1:20 dilution in media.Count the cells in the dilution using a hemocytometer. Then, dilute the main suspension as needed, using media with rock inhibitor. To proceed, add the required number of cells onto each prepared stencil.Only a mono-layer of cells should be deposited, so spread them out evenly. Let the stencil sit undisturbed for five minutes, so the cells can settle. Then, carefully transfer the loaded stencils to a incubator, keeping the dishes level throughout the transfer.Let the cells attach to the stencils over an hour-long incubation. After the cells have been allowed to attach, examine them under a microscope to ensure that they did so properly. Transfer the stencil back to the hood, and aspirate the unattached cells off the stencil.Next, add two milliliters of 0.5%compatible non-ionic surfactant in DMF12 to the area surrounding the stencil. Swirl the solution around to cover the stencil. Then, use a pair of autoclaved forceps to gently peel the stencil off the dish.After removing the stencil, inspect the micropatterend cells under a microscope. The non-ionic surfactant prevents these cells from drying out. Let the micropatterend cells incubate in the surfactant-supplemented media solution for 10 minutes at 37 degrees Celsius.After 10 minutes, aspirate away the solution, and wash the plate with two milliliters of normal DMF12 medium, three times. After the washes, add two milliliters of HESC maintenance medium with rock inhibitor. Once again, inspect the cells to check that the micropatterns have formed properly.Then, incubate the cells at 37 degrees Celsius overnight. The next day, aspirate the medium, wash the cells once with DMF12, and then, add two milliliters of supplemented differentiation medium, to induce the cells to undergo mesoendoderm differentiation. Later, analyze the results.The fabrication of the cell stencil using a laser cutter, provided 1000 micron features. The stencil was composed of a thin stenciling sheet with micropatterned through holes, and a PDMS gasket used to contain the cells. Using this tool, micropatterened HESCs were investigated during mesoendoderm differentiation.H9-HESC colony peripheries have higher integrin adhesions than cells at the colony interior. Thus, the cells differentiate into brachiary T-positive mesoendoderm cells, when induced with activan, BMP4, and FGF2. When such cells were patterned into monolayers of different geometries, geometric and isotropiate right angles and at convex curvatures, led to a local concentration of integrin mediated traction forces, and resulted in increased mesoendoderm differentiation.T expression intensity at different localities within a single colony showed that the extent of meso endoderm induction was indeed higher in sharp corners of square and rectangular colonies, and at the convex curvatures of a semi-circular arc. This demonstrates that cell adhesion mediated mechanical forces can be modulated using the described method. Before attempting the procedure, it is important to optimize the matrix coating time, the cell digestion and attachment time, for the specific human pluripotent stem cell line that you are working with.

Research

JoVE Journal

Developmental Biology

Large-Scale Production of Cardiomyocytes from Human Pluripotent Stem Cells Using a Highly Reproducible Small Molecule-Based Differentiation Protocol

Die Großproduktion von Kardiomyozyten aus humanen pluripotenten Stammzellen hochreproduzierbarer Kleinmolekül-basierte Differenzierung Protokoll

Maximizing the benefit of human pluripotent stem cells (hPSCs) for research, disease modeling, pharmaceutical and clinical applications requires robust ...

Forschung

Entwicklungsbiologie

Aggregate Size Optimization in Microwells for Suspension-based Cardiac Differentiation of Human Pluripotent Stem Cells

Aggregate Größenoptimierung in Mikrotiterplattenvertiefungen für Abhänge-basierte Herz Differenzierung von humanen pluripotenten Stammzellen

Cardiac differentiation of human pluripotent stems cells (hPSCs) is typically carried out in suspension cell aggregates.  Conventional aggregate formation ...

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

Schnelle Neuronale Differenzierung von induzierten pluripotenten Stammzellen zur Messung der Netzwerkaktivität auf Micro-Elektroden-Arrays

Neurons derived from human induced Pluripotent Stem Cells (hiPSCs) provide a promising new tool for studying neurological disorders. In the past decade, ...

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

Differenzierung der Primitive und Definitive hämatopoetischen Vorläuferzellen aus menschlichen pluripotente Stammzellen gerichtet

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

Efficient Differentiation of Pluripotent Stem Cells to NKX6-1+ Pancreatic Progenitors

Efficient Differentiation of Pluripotent Stem Cells to NKX6-1+ Pancreatic Progenitors

Effiziente Differenzierung von pluripotenten Stammzellen zu NKX6-1<sup&gt; +</sup&gt; Pancreatic Vorläufern

Pluripotent stem cells have the ability to self renew and differentiate to multiple lineages, making them an attractive source for the generation of ...

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

In vitro Differenzierung von humanen pluripotenten Zellen in Trophoblastische Stammzellen

The placenta is the first organ to develop during embryogenesis and is required for the survival of the developing embryo. The placenta is comprised of ...

Rapid, Directed Differentiation of Retinal Pigment Epithelial Cells from Human Embryonic or Induced Pluripotent Stem Cells

Schnelle, gezielte Differenzierung der retinalen Pigment epithelialen Zellen aus menschlichen embryonalen oder induzierte pluripotente Stammzellen

We describe a robust method to direct the differentiation of pluripotent stem cells into retinal pigment epithelial cells (RPE). The purpose of providing ...

Efficient Differentiation of Human Pluripotent Stem Cells into Liver Cells

Effiziente Differenzierung menschlicher pluripotenter Stammzellen in Leberzellen

The liver detoxifies harmful substances, secretes vital proteins, and executes key metabolic activities, thus sustaining life. Consequently, liver ...

In Vitro Generation of Somite Derivatives from Human Induced Pluripotent Stem Cells

In vitro-Generation der Somiten Derivate aus menschlichen induzierten Zellen pluripotenten Stammzellen

In response to signals such as WNTs, bone morphogenetic proteins (BMPs), and sonic hedgehog (SHH) secreted from surrounding tissues, somites (SMs) give ...

Mapping the Emergent Spatial Organization of Mammalian Cells using Micropatterns and Quantitative Imaging

Kartierung der Emergent Spatial Organization of Mammalian Cells mit Mikromustern und quantitativer Bildgebung

A fundamental goal in biology is to understand how patterns emerge during development. Several groups have shown that patterning can be achieved in vitro ...