Name: generation of ipsc-derived human brain organoids to model early neurodevelopmental disorders
Uploaded: 2017-04-14T13:00:00
Description: Watch this Scientific Journal Video about Generation of iPSC-derived Human Brain Organoids to Model Early Neurodevelopmental Disorders at JoVE.com

Diese Arbeit wurde von der Fritz-Thyssen-Stiftung (Az.10.14.2.152) unterstützt. Wir sind dankbar, dass die Gewebeeinbettung Anlage und das Mikroskop Core Facility des ZMMK. Wir sind dankbar für die Diskussionen und technische von den Mitgliedern des Labors für Zentrosom und Zytoskelett Biologie unterstützt. Wir danken Li Ming Gooi das Manuskript für das Korrekturlesen.

1. Erzeugung von Brain Organoide (23 Tage) <ol><li> Initiierung von Neuroektoderms (5 Tage) HINWEIS: Die folgenden Punkte sollten vor dem Beginn der Differenzierung in Betracht gezogen werden. Die Umprogrammierung Methode (lentiviral-, Sendai-Virus-, Episomen oder microRNA-Basis etc.) menschlichen iPSCs erhalten sollte idealerweise die gleiche für alle Patienten und iPSC Leitungen steuern. Verschiedene Umprogrammierung Kits und Anweisungen auf der Grundlage veröffentlichter Protokoll...

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MCPH ist eine komplexe menschliche Erkrankung des Nervensystems , die nicht in Tiermodellen in vivo oder in einfachen menschlichen Zellkulturansätze in vitro rekapituliert werden kann. Die klinische Manifestation der MCPH beginnt während des ersten Trimesters zu erscheinen, wenn sie früh beginnt die Neurogenese. Somit stellen die 3D Gehirn Organoide ein zuverlässiges experimentelles System MCPH Entwicklung zu modellieren. Darüber hinaus 3D menschliches Gehirn Organoide ist ein idealer Ansatz, da i...

Menschliche Entwicklung des Nervenstörungen, wie Mikrozephalie, nur schlecht in Tiermodellen untersucht werden, aufgrund der Tatsache, dass das menschliche Gehirn haben eine erweiterte kortikalen Oberfläche, ein einzigartiges Merkmal der sich von nicht-menschlichen Tieren. Dieser Aspekt macht die menschliche Entwicklung des Gehirns einen komplexen Prozess, der nicht ausreichend in einem 2D untersucht werden, in vitro Zellkultursystem. Schwellen 3D-Kulturtechniken ermöglichen die Erzeugung von gewebeähnlichen Organoide aus induzierten pluripotenten Stammzellen (iPS). Die in - vitro - Differenzierung von pluripotenten Stammzellen in einer 3D - Suspensionskultur ermöglicht die Bildung von verschiedenen Zelltypen in einer angemessenen und regionsspezifische Weise, was zu einem organisierten, geschichteten Gewebe 1, 2, 3. Dank Laboratorien, die 3D-Kulturtechnologien Pionierarbeit geleistet und die Komplexität der Organbildung entmystifiziert, aus Stammzellen beginnen,entwickeln wir eine robuste Methode Gehirn Organoide der Erzeugung frühe Ereignisse der menschlichen Gehirn Entwicklung zu beschreiben und Mikrozephalie in vitro 1, 2, 3 zu modellieren. Es ist bemerkenswert , dass wir die ursprüngliche Methode , entwickelt von Lancaster et al angepasst. zerebrale Organoide 1 zu erzeugen. Diese Methode wurde nach unseren experimentellen Anforderungen modifiziert. Das Ziel einer Studie von Gabriel et al. war die zellulären und molekularen Mechanismen der neuronalen Stammzell Wartung während der Entwicklung des Gehirns zu untersuchen. Um dies zu tun, wurde eine mechanistische Studie durch die Analyse von neuralen Vorläuferzellen (NSC) in 3D durchgeführt Gehirn Organoide von einem Patienten stammte Mikrocephalie 4. Dieser Patient trug eine Mutation in CPAP, ein konserviertes Protein zentrosomalen für Zentrosom Biogenese 5 erforderlich. Eine weithin akzeptierte hypoThese ist , dass Mikrocephalie das Ergebnis einer Erschöpfung des NPC - Pools ist, und dies könnte entweder zum Zelltod oder zu einem vorzeitigen Differenzierung 1, 6, 7, 8, 9 liegt. Durch die Analyse der ventrikulären Zonen (VZs) von Mikrocephalie brain Organoiden, wurde gezeigt , dass eine beträchtliche Anzahl von NSC asymmetrischer Zellteilung zu unterziehen, im Gegensatz zu Gehirn Organoide von einem gesunden Spender stamm 4. Umfangreiche mikroskopische und biochemische Analysen von microcephalic Gehirn Organoide ergab eine unerwartete Rolle für CPAP in rechtzeitige Zilien Demontage 4. Insbesondere mutierte CPAP mit retardierten cilium Demontage- und verzögerte Zellzyklus - Wiedereintritt verbunden ist , auf die Differenzierung von vorzeitiger NSC führenden 4. Diese Ergebnisse legen nahe, eine Rolle für Zilien in Mikrozephalie und thEIR Beteiligung während der Neurogenese und Gehirngröße Kontrolle 10. Der erste Teil dieses Protokolls ist eine Beschreibung eines dreistufigen Verfahrens homogene brain Organoiden zu erzeugen. Wie bereits erwähnt, wurde das ursprüngliche Lancaster Protokoll angepasst und modifiziert unser Ziel 1 zu entsprechen. Erstens sind menschliche iPSCs Kultur in einem definierten feeder freien Zustand auf Engelbreth-Holm-Swarm (EHS) Matrix. Dieser Schritt vermeidet die Variationen von Feeder-abhängigen pluripotenten Stammzellkulturen. In diesem Protokoll zur Induktion neuronaler Differenzierung zu bilden direkt von iPSCs neuralen Epithel beginnt. Durch den Embryoidkörper (EB) Bildungsschritt übersprungen, die neurale Differenzierung erfolgt in einer kontrollierteren und Weise gerichtet. Dieser Ansatz begrenzt die spontane und ungerichteten Bildung anderer Keimzellschichten, wie Mesoderm und Endoderm. Durch dieses Protokoll anwenden, können Neurosphären neuronale Rosetten enthalten am Tag geerntet werden 5 für EHS-Matrix und Einbettung stationäre Suspensionskultur. Das organoiden Medium für den dritten Schritt unseres Protokolls verwendet wird mit Dorsomorphin und SB431542 ergänzt. Dorsomorphin ist ein niedermolekularer Inhibitor von knochenmorphogenes Protein (BMP) und SB431542 hemmt die TGF &amp; bgr; / Activin / Knoten Signalweg. Die Kombination dieser Faktoren könnte neuronale Differenzierung mehr fördert effizienter als Retinsäure allein 11, 12, 13, 14. Insgesamt ermöglichen diese Modifikationen die reproduzierbare Erzeugung von brain Organoide mit minimalen Variationen in Organoiden. Wichtig ist, dass diese Methode microcephalic Gehirn Organoide von Patienten zu iPSCs robust erzeugt aufgebracht, die Mutationen in Genen tragen, die Zentrosomen und Zellzyklusdynamik beeinflussen. Der zweite Teil dieses Protokolls gibt Anweisungen br vorzubereitenain Organoide für die Analyse und Interpretation von zellulären Defekte in Mikrozephalie. Dazu gehören Fixierung, Kryoschneiden, Immunofluoreszenzfärbung und konfokale mikroskopische Analyse. Dieses Protokoll wird dem Leser eine detaillierte Beschreibung der erwarteten Ergebnisse liefern und mit Anleitung zur Interpretation.

<table border="0" cellpadding="0" cellspacing="0" width="670">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Anti-mouse 488</td>
			<td style="width:123px;">Invitrogen</td>
			<td style="width:119px;">A-11001</td>
			<td style="width:212px;">Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 488</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Anti-rabbit 647</td>
			<td style="width:123px;">Invitrogen</td>
			<td style="width:119px;">A-21245</td>
			<td style="width:212px;">Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 647</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Arl13b</td>
			<td style="width:123px;">proteintech</td>
			<td style="width:119px;">17711-1-AP</td>
			<td style="width:212px;">ARL13B rabbit polyclonal antibody&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">CELLSPIN system</td>
			<td style="width:123px;">IBS Integra Bioscience</td>
			<td style="width:119px;">183001</td>
			<td style="width:212px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">DAPI</td>
			<td style="width:123px;">Sigma-Aldrich, US</td>
			<td style="width:119px;">32670</td>
			<td style="width:212px;">4&#8242;,6-Diamidino-2-phenylindole dihydrochloride; multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">DMEM/F-12</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">31331093</td>
			<td style="width:212px;">Dulbecco&#39;s Modified Eagle Medium: Nutrient Mixture F-12&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dorsomorphin</td>
			<td style="width:123px;">Sigma-Aldrich, US</td>
			<td style="width:119px;">P5499</td>
			<td style="width:212px;">Compound C; multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Embedding medium</td>
			<td style="width:123px;">AppliChem</td>
			<td style="width:119px;">A9011, 0100</td>
			<td style="width:212px;">Mowiol; embedding medium; multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Engelbreth-Holm-Swarm (EHS) matrix</td>
			<td style="width:123px;">Corning</td>
			<td style="width:119px;">354277</td>
			<td style="width:212px;">Matrigel hESC-qualified matrix; important: hESC qualified</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Fish gelatin&#160;</td>
			<td style="width:123px;">Sigma-Aldrich, US</td>
			<td style="width:119px;">G7765-250ML</td>
			<td style="width:212px;">Gelatin from cold water fish skin; multiple suppliers; autoclave after adding to PBS to dissolve and sterilize, store at 4&#176;C</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Glycine</td>
			<td style="width:123px;">AppliChem</td>
			<td style="width:119px;">A1067,1000</td>
			<td style="width:212px;">Glycine for molecular biology; multiple suppliers&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Inoculation loop with needle, disposable (1 &#181;l)</td>
			<td style="width:123px;">Sigma Aldrich, US</td>
			<td style="width:119px;">BR452201-1000EA</td>
			<td style="width:212px;">multiple suppliers&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Insulin</td>
			<td style="width:123px;">Sigma-Aldrich, US</td>
			<td style="width:119px;">I3536-100MG</td>
			<td style="width:212px;">multiple suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">L-glutamine</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">25030081</td>
			<td style="width:212px;">L-glutamine (200 mM)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Medium A</td>
			<td style="width:123px;">Stem cell technologies</td>
			<td style="width:119px;">#05850</td>
			<td style="width:212px;">mTeSR1 (hiPSC medium)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Medium B</td>
			<td style="width:123px;">Stem cell technologies</td>
			<td style="width:119px;">#05835</td>
			<td style="width:212px;">Neural induction medium (NIM); neural differentiation medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Medium C</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">21103049</td>
			<td style="width:212px;">Neural Basal Medium</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">MEM</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">11140035</td>
			<td style="width:212px;">MEM non-essential amino acids solution (100x)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">MycoAlert Mycoplasma Detection Kit</td>
			<td style="width:123px;">Lonza, Switzerland</td>
			<td style="width:119px;">#LT07-218</td>
			<td style="width:212px;">Mycoplasma detection kit; multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Nestin</td>
			<td style="width:123px;">Novus biologicals</td>
			<td style="width:119px;">NBP1-92717</td>
			<td style="width:212px;">Nestin mouse monoclonal antibody (4D11)</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Paraformaldehyde (PFA)</td>
			<td style="width:123px;">AppliChem</td>
			<td style="width:119px;">A3813, 0500</td>
			<td style="width:212px;">4% in PBS, store solution at -20&#176;C; caution: wear skin and eye protection and work under hood&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">PBS tablets</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">18912014</td>
			<td style="width:212px;">See manufacturer&#180;s instructions; multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Penicillin-Streptomycin (10.000 U/ml)</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">15140122</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Poly-L-lysine solution (PLL)</td>
			<td style="width:123px;">Sigma-Aldrich, US</td>
			<td style="width:119px;">P8920-100ML</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">pVim</td>
			<td style="width:123px;">MBL</td>
			<td style="width:119px;">D076-3S</td>
			<td style="width:212px;">Phospho-Vimentin (Ser55) mAb</td>
		</tr>
		<tr height="147">
			<td height="147" style="height:147px;width:216px;">Reagent A&#160;</td>
			<td style="width:123px;">Stem cell technologies</td>
			<td style="width:119px;"># 05872</td>
			<td style="width:212px;">Note to Protocol 1.1.1.2; ReLSR (Enzyme-free human ES and iPS cell selection and passaging reagent); please follow manufactorer&#180;s protocol; alternative products from muliple suppliers available</td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:216px;">Reagent B&#160;</td>
			<td style="width:123px;">Sigma-Aldrich, US</td>
			<td style="width:119px;">A6964-100ML</td>
			<td style="width:212px;">Accutase solution is an enzymatic solution for single cell dissociation; multiple suppliers; protocol 1.1.2 &#34;enzymatic cell dissociation solution&#8221;&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Research Cryostat Leica CM3050 S</td>
			<td style="width:123px;">Leica biosystems</td>
			<td style="width:119px;">CM3050 S</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">SB431542</td>
			<td style="width:123px;">Selleckchem.com</td>
			<td style="width:119px;">S1067</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Spinner flask 250 ml</td>
			<td style="width:123px;">IBS Integra Bioscience</td>
			<td style="width:119px;">182026</td>
			<td style="width:212px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sucrose</td>
			<td style="width:123px;">AppliChem</td>
			<td style="width:119px;">A4734, 1000</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Superfrost ultra plus microscope slides</td>
			<td style="width:123px;">Thermo scientific, US</td>
			<td style="width:119px;">J3800AMNZ</td>
			<td style="width:212px;">Slides should be labeled with a &#34;+&#34; and positively charged</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Supplement 1</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">17502048</td>
			<td style="width:212px;">N-2 supplement (100x)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Supplement 2 w/o Vitamin A</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">12587010</td>
			<td style="width:212px;">B-27 supplement (50x), minus vitamin A; multiple suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tissue-Tek Cryomold</td>
			<td style="width:123px;">Sakura, NL</td>
			<td style="width:119px;">4565</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tissue-Tek O.C.T. compound</td>
			<td style="width:123px;">Sakura, NL</td>
			<td style="width:119px;">4583</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Triton X-100</td>
			<td style="width:123px;">AppliChem</td>
			<td style="width:119px;">A1388,0500</td>
			<td style="width:212px;">Multiple suppliers multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">TUJ1</td>
			<td style="width:123px;">Sigma-Aldrich, US</td>
			<td style="width:119px;">T2200</td>
			<td style="width:212px;">&#946;-Tubulin III (rabbit polyclonal)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">TUNEL assay</td>
			<td style="width:123px;">Promega, US</td>
			<td style="width:119px;">G3250</td>
			<td style="width:212px;">DeadEnd Fluorometric TUNEL system; multiple suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tween 20 for molecular biology</td>
			<td style="width:123px;">AppliChem</td>
			<td style="width:119px;">A4974,0500</td>
			<td style="width:212px;">Multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">waterproof sheet</td>
			<td style="width:123px;">BEMIS company, inc.</td>
			<td style="width:119px;">PM996</td>
			<td style="width:212px;">Parafilm &#8220;M&#8221;; multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Y-27632&#160;</td>
			<td style="width:123px;">Selleckchem.com</td>
			<td style="width:119px;">S1049</td>
			<td style="width:212px;">ROCK-inhibitor (Y-27632 2HCL); multiple suppliers</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">&#946;-mercaptoethanol</td>
			<td style="width:123px;">Gibco, US</td>
			<td style="width:119px;">31350010</td>
			<td style="width:212px;">2-mercaptoethanol (50 mM); multiple suppliers</td>
		</tr>
	</tbody>
</table>

generation of ipsc-derived human brain organoids to model early neurodevelopmental disorders

The restricted availability of suitable in vitro models that can reliably represent complex human brain development is a significant bottleneck that limits the translation of basic brain research into clinical application. While induced pluripotent stem cells (iPSCs) have replaced the ethically questionable human embryonic stem cells, iPSC-based neuronal differentiation studies remain descriptive at the cellular level but fail to adequately provide the details that could be derived from a complex, 3D human brain tissue. 
This gap is now filled through the application of iPSC-derived, 3D brain organoids, "Brains in a dish," that model many features of complex human brain development. Here, a method for generating iPSC-derived, 3D brain organoids is described. The organoids can help with modeling autosomal recessive primary microcephaly (MCPH), a rare human neurodevelopmental disorder. A widely accepted explanation for the brain malformation in MCPH is a depletion of the neural stem cell pool during the early stages of human brain development, a developmental defect that is difficult to recreate or prove in vitro. 
To study MCPH, we generated iPSCs from patient-derived fibroblasts carrying a mutation in the centrosomal protein CPAP. By analyzing the ventricular zone of microcephaly 3D brain organoids, we showed the premature differentiation of neural progenitors. These 3D brain organoids are a powerful in vitro system that will be instrumental in modeling congenital brain disorders induced by neurotoxic chemicals, neurotrophic viral infections, or inherited genetic mutations.

Die Erzeugung von brain Organoide erfordert mindestens drei Wochen ununterbrochener Kultivierung (1A). Um reproduzierbare Ergebnisse zu erreichen, empfehlen wir, dass die Forscher jeden Schritt dokumentiert und, was wichtig ist, vermeidet jegliche Änderungen in Bezug auf Medienkomponenten, Zeitpunkte und die Handhabung von Zellen. Hier geben wir einen Überblick darüber, wie zu beurteilen, ob kritische Meilensteine ​​erreicht, um sich Organoide von ausreichender Qu...

Watch this Scientific Journal Video about Generation of iPSC-derived Human Brain Organoids to Model Early Neurodevelopmental Disorders at JoVE.com

Generation of iPSC-derived Human Brain Organoids to Model Early Neurodevelopmental Disorders

Modeling human brain development has been hindered due to the unprecedented complexity of neural epithelial tissue. Here, a method for the robust generation of brain organoids to delineate early events of human brain development and to model microcephaly in vitro is described.

Die Erzeugung von iPS-derived Human Brain Organoide auf Modell Frühe Störungen der neurologischen Entwicklung

The restricted availability of suitable in vitro models that can reliably represent complex human brain development is a significant bottleneck that ...

Research

JoVE Journal

Developmental Biology

Rapid Detection of Neurodevelopmental Phenotypes in Human Neural Precursor Cells (NPCs)

Rapid Detection of Neurodevelopmental Phenotypes in Human Neural Precursor Cells NPCs

Schnelle Erkennung von Entwicklungsstörungen Phänotypen in menschlichen neuronalen Vorläuferzellen (NPCs)

Human brain development proceeds through a series of precisely orchestrated processes, with earlier stages distinguished by proliferation, migration, and ...

Forschung

Entwicklungsbiologie

Generation of Standardized and Reproducible Forebrain-type Cerebral Organoids from Human Induced Pluripotent Stem Cells

Generation von standardisierten und reproduzierbaren Vorderhirn-Typ zerebralen Organellen aus menschlichen induzierten pluripotenten Stammzellen

The human cortex is highly expanded and exhibits a complex structure with specific functional areas, providing higher brain function, such as cognition. ...

Zika Virus Infection of Cultured Human Fetal Brain Neural Stem Cells for Immunocytochemical Analysis

Zika-Virus-Infektion der kultivierten menschlichen fetalen Gehirns neurale Stammzellen für Immunocytochemical Analyse

Human fetal brain neural stem cells are a unique non-genetically modified model system to study the impact of various stimuli on human developmental ...

Defined Xeno-free and Feeder-free Culture Conditions for the Generation of Human iPSC-derived Retinal Cell Models

Definierten Xeno-frei und Feeder-freie Kulturbedingungen für die Generation der menschlichen iPSC-abgeleitete Retinal Zellmodellen

The production of specialized cells from pluripotent stem cells provides a powerful tool to develop new approaches for regenerative medicine. The use of ...

An In Vitro 3D Model and Computational Pipeline to Quantify the Vasculogenic Potential of iPSC-Derived Endothelial Progenitors

Ein In-Vitro-3D-Modell und eine Computational Pipeline zur Quantifizierung des vaskulogenen Potenzials von iPSC-derived endotheliaalen Vorläufern

Induced pluripotent stem cells (iPSCs) are a patient-specific, proliferative cell source that can differentiate into any somatic cell type. Bipotent ...

Generation of Organoids from Mouse Extrahepatic Bile Ducts

Generation von Organellen von Maus extrahepatische Gallenwege

Cholangiopathies, which affect extrahepatic bile ducts (EHBDs), include biliary atresia, primary sclerosing cholangitis, and cholangiocarcinoma. They have ...

Generation of Induced Neural Stem Cells from Peripheral Mononuclear Cells and Differentiation Toward Dopaminergic Neuron Precursors for Transplantation Studies

Generierung induzierter neuronaler Stammzellen aus peripheren mononukleären Zellen und Differenzierung zu dopaminergen Neuronenvorstufen für Transplantationsstudien

Parkinson&#39;s disease (PD) is caused by degeneration of dopaminergic (DA) neurons at the substantia nigra pars compacta (SNpc) in the ventral ...

A Static Self-Directed Method for Generating Brain Organoids from Human Embryonic Stem Cells

Eine statische selbstgesteuerte Methode zur Generierung von Hirnorganoiden aus menschlichen embryonalen Stammzellen

Human brain organoids differentiated from embryonic stem cells offer the unique opportunity to study complicated interactions of multiple cell types in a ...

Generation of Multicellular Human Primary Endometrial Organoids

Generierung von mehrzelligen humanen primären Endometrium-Organoiden

The human endometrium is one of the most hormonally responsive tissues in the body and is essential for the establishment of pregnancy. This tissue can ...

Generation of Mosaic Mammary Organoids by Differential Trypsinization

Generierung von Mosaik-Mammary-Organoiden durch Differential-Trypsinisierung

Organoids offer self-organizing, three-dimensional tissue structures that recapitulate physiological processes in the convenience of a dish. The murine ...