Name: generation of human neurons and oligodendrocytes from pluripotent stem cells for modeling neuron-oligodendrocyte interactions
Uploaded: 2020-11-09T15:00:00
Description: Watch this Scientific Journal Video about Generation of Human Neurons and Oligodendrocytes from Pluripotent Stem Cells for Modeling Neuron-Oligodendrocyte Interactions at JoVE.com

Este trabajo fue apoyado por las subvenciones de los Institutos Nacionales de Salud (R00 AG054616 a Y.A.H. y T32 GM136566 a K.C.), la Escuela de Medicina de la Universidad de Stanford y una beca Siebel (otorgada a SC). Y.A.H. es profesor traslacional de GFL del Centro de Neurociencia Traslacional del Instituto Brown de Ciencias Traslacionales.

1. Inducción de neuronas humanas a partir de células madre pluripotentes humanas
<ol><li>Preparación de lentivirus (~ 5 días, protocolo detallado como se describió anteriormente16)<ol><li>Placa ~1 millón de células HEK293T cada matraz T75, para tenerlas ~40% confluentes al realizar la transfección. Transfectarlos con plásmidos que expresan Ngn2 inducible por tetraciclina y gen resistente a la puromicina (PuroR; bajo el mismo control del promotor TetO), rtTA y los tres plásmidos auxilia...

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H., Park, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+reprogramming+methods+for+generation+of+induced-oligodendrocyte+precursor+cells.">Comparison of reprogramming methods for generation of induced-oligodendrocyte precursor cells.</a> Biomolecules &amp; Therapeutics (Seoul). 25 (4), 362-366 (2017).</li><li>Ehrlich, 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=Rapid+and+efficient+generation+of+oligodendrocytes+from+human+induced+pluripotent+stem+cells+using+transcription+factors.">Rapid and efficient generation of oligodendrocytes from human induced pluripotent stem cells using transcription factors.</a> Proceedings of the National Academy of Sciences of the United States of America. 114 (11), 2243-2252 (2017).</li><li>Rodrigues, G. M. 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Además del apoyo físico y metabólico para estabilizar las estructuras sinapsis y facilitar la conducción de señales saltatorias por mielinización, las células del linaje de oligodendrocitos pueden dar forma al patrón de actividad neuronal a través de conversaciones cruzadas rápidas y dinámicas con las neuronas 5,6,7. Mientras que en la patología de la EA las respuestas oligodendrogliales se consideraron inicialmente ...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/61778/generation-human-neurons-oligodendrocytes-from-pluripotent-stem-cells">Generation of Human Neurons and Oligodendrocytes from Pluripotent Stem Cells for Modeling Neuron-Oligodendrocyte Interactions</a>.&#160;The Representative Results section has been updated.
Figure 3 was updated from:
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/61778/61778fig03.jpg" /> 
Figure 3: Co-culture of iNs and iOPCs.&#160;(A) Representative bright field image of co-cultured iNs and iOPCs at Day 7, showing a proper density for further maturation. (B) Representative immunofluorescence image of iNs and iOPCs co-cultured for 28 days. Axonal marker neurofilament NF is shown in green and oligodendrocytic marker MBP in red. Right, a segment of iN axon ensheathed by iOL process (MBP+). (C) Synapse formation assayed in 4-week-old co-cultures. Cells were stained for Synapsin 1 (Syn1, green) and MAP2 (red), and synaptic puncta were quantified by confocal analysis of density along the dendritic segments as described17,18. (D) In our co-cultures of iNs and iOPCs (7 days of co-culturing), the expression of astrocyte markers, ALDHL1 and GFAP, is minimal (top), and the expression of microglia markers, TMEM119, TREM2, and CD33, is not detected (N.D.) by qPCR. The contamination from these two glial cell types is thus excluded. <a href="https://www.jove.com/files/ftp_upload/61778/61778fig03large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
to:
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/61778/61778fig3v2.png" /> 
Figure 3: Co-culture of iNs and iOPCs.&#160;(A) Representative bright field image of co-cultured iNs and iOPCs at Day 7, showing a proper density for further maturation. (B) Representative immunofluorescence image of iNs and iOPCs co-cultured for 28 days. Axonal marker neurofilament NF is shown in green and oligodendrocytic marker MBP in red. Right, a segment of iN axon ensheathed by iOL process (MBP+). (C) Synapse formation assayed in 4-week-old co-cultures. Cells were stained for Synapsin 1 (Syn1, green) and MAP2 (red), and synaptic puncta were quantified by confocal analysis of density along the dendritic segments as described17,18. (D) In our co-cultures of iNs and iOPCs (7 days of co-culturing), the expression of astrocyte markers, ALDHL1 and GFAP, is minimal (top), and the expression of microglia markers, TMEM119, TREM2, and CD33, is not detected (N.D.) by qPCR. The contamination from these two glial cell types is thus excluded.&#160;(E) Coculturing iOPC with iN leads to the formation of neuron-OPC synapses. The fluorescence-tagged post-synaptic marker PSD95-mCherry is expressed only in OPCs, and display a diffuse pattern in single cultures (left) but aggregate to form puncta in cocultures (right, indicated by arrows; Tuj1, neuronal marker). (F) The expression of well-characterized oligodendroglial genes that can sense and respond to neuronal activities in the pure cultures of iOPCs at Day 14.&#160;<a href="https://www.jove.com/files/ftp_upload/61778/61778fig3v2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
The fourth&#160;paragraph was updated from:
Co-culturing of iNs and iOPCs 
This protocol is optimized specifically for co-culturing iNs and iOPCs and allow our real-time monitoring of the inter-cellular communications between these two cell types along the course of neural development. The ideal plating densities for both cell types need to be decided with a series of cell number titration to achieve proper differentiation (Figure 3A). After 4 weeks in co-cultures, the iOPCs are expected to be adequately differentiated into OLs that are positive for specific markers such as MBP and extend processes to ensheath axons (Figure 3B). The co-culture system can robustly boost up the number of synapses, indicating that the iOPCs provide a neuronal support through physical contacts or release of trophic factors (Figure 3C). We can maintain the co-cultures in acceptable health condition for up to 6 weeks and observe that the synapse number and other neuronal attributes plateau around the fifth week. Of note, astrocytes and microglia are not present in our preparations and their absence can be documented by checking the expression of specific markers (Figure 3D).
to:
Co-culturing of iNs and iOPCs 
This protocol is optimized specifically for co-culturing iNs and iOPCs and allow our real-time monitoring of the inter-cellular communications between these two cell types along the course of neural development. The ideal plating densities for both cell types need to be decided with a series of cell number titration to achieve proper differentiation (Figure 3A). After 4 weeks in co-cultures, the iOPCs are expected to be adequately differentiated into OLs that are positive for specific markers such as MBP and extend processes to ensheath axons (Figure 3B). The co-culture system can robustly boost up the number of synapses, indicating that the iOPCs provide a neuronal support through physical contacts or release of trophic factors (Figure 3C). We can maintain the co-cultures in acceptable health condition for up to 6 weeks and observe that the synapse number and other neuronal attributes plateau around the fifth week. Of note, astrocytes and microglia are not present in our preparations and their absence can be documented by checking the expression of specific markers (Figure 3D).&#160;The iOPCs express a good number of well-characterized genes that can potentially respond to and mediate the activity-dependent signals from neighboring neurons, in a paracrine (e.g. neurotrophins and metabolites) and/or a synaptic manner (Figure&#160;3E and 3F).&#160;

An erratum was issued for:&#160;<a href="https://www.jove.com/t/61778/generation-human-neurons-oligodendrocytes-from-pluripotent-stem-cells">Generation of Human Neurons and Oligodendrocytes from Pluripotent Stem Cells for Modeling Neuron-Oligodendrocyte Interactions</a>.&#160;The Representative Results section has been updated.

Erratum: Generation of Human Neurons and Oligodendrocytes from Pluripotent Stem Cells for Modeling Neuron-Oligodendrocyte Interactions

Las células del linaje de oligodendrocitos, incluidas las células precursoras de oligodendrocitos (OPC), los oligodendrocitos mielinizantes y los tipos de transición intermedios, constituyen un grupo importante de células cerebrales humanas1 que participan activamente en muchas funciones críticas para el correcto funcionamiento y mantenimiento de nuestro sistema nervioso central a lo largo del desarrollo neuronal y el envejecimiento 2,3,4 . Mientras que los oligodendrocitos son bien conocidos por producir mielina para facilitar la transmisión de la actividad neuronal y apoyar la salud axonal en la sustancia blanca, los OPC son abundantes (~ 5%) en la materia gris donde la mielinización es escasa y realizan funciones de señalización dependientes de la actividad para gobernar el comportamiento de aprendizaje y la formación de la memoria 5,6,7,8 . La forma en que las células oligodendrogliales funcionan y la disfunción en la patogénesis de la enfermedad de Alzheimer (EA) y otras afecciones neurodegenerativas asociadas a la edad han sido poco estudiadas9. Las insuficiencias de un sistema modelo apropiado y las deficiencias en el conocimiento general para guiar un camino experimental hacia adelante son las principales razones de esta brecha.
A la luz de los últimos avances en la obtención de células cerebrales humanas a partir de células madre pluripotentes, incluidas las células madre embrionarias (ES) y las células madre pluripotentes inducidas (iPS), tales modelos celulares junto con las herramientas modernas de edición de genes han surgido como herramientas robustas para manejar el intrincado nexo de las interacciones celulares en el cerebro, y son capaces de demostrar manifestaciones de enfermedades específicas del ser humano10, 11. Teniendo en cuenta que los tipos individuales de células cerebrales pueden exhibir efectos distintos e incluso conflictivos frente a las mismas condiciones promotoras de la EA12,13, esta metodología de células madre ofrece de manera única información específica del tipo de célula que anteriormente se había pasado por alto utilizando modelos establecidos in vivo o in vitro que solo proporcionan lecturas agregadas de colecciones de tipos de células cerebrales. En la última década, se han desarrollado un buen número de protocolos confiables para generar neuronas humanas a partir de la transdiferenciación de células ES/iPS o la conversión directa de otros tipos de células diferenciadas terminalmente (por ejemplo, fibroblastos)14,15. En particular, la aplicación de factores de transcripción neurogénicos clave (por ejemplo, neurogenina 2, Ngn2)16 a células madre pluripotentes humanas puede generar una población homogénea de tipos de células neuronales bien caracterizadas para cultivos puros sin necesidad de cocultivo con células gliales12,17,18. Para los oligodendrocitos humanos inducidos, hay algunos protocolos publicados que pueden generar células funcionales muy parecidas a sus contrapartes primarias, con una amplia gama de eficiencia y demanda en tiempo y recursos 19,20,21,22,23,24,25,26,27,28 . Hasta la fecha, ninguno de estos protocolos se ha aplicado para investigar cómo las células oligodendrogliales responden y afectan la patogénesis de la EA.
Aquí, describimos nuestros protocolos mejorados para cultivos únicos y mixtos de neuronas inducidas humanas (iNs) y OPC / oligodendrocitos (iOPC / iOL). El protocolo iN descrito aquí se basa en el enfoqueNgn2 16 ampliamente utilizado, y tiene la característica adicional de estar libre de glía. Los iNs resultantes son homogéneos y se asemejan mucho a las neuronas excitadoras de la capa cortical 2/3, con morfología piramidal característica, patrón de expresión génica y características electrofisiológicas17,18 (Figura 1). Para superar algunas de las barreras fundamentales en la diferenciación dirigida de células madre pluripotentes, hemos desarrollado un método simple y eficaz de pretratamiento con dosis bajas de dimetilsulfóxido (DMSO)29,30, y hemos reportado una mayor propensión de las células ES/iPS humanas a transdiferenciarse en iOPCs e iOLs31, basado en un protocolo ampliamente adaptado por Douvaras y Fossati 32 . Hemos simplificado aún más el protocolo e incorporado un compuesto robusto que promueve la diferenciación, clemastine 7,33,34, para acelerar el proceso de maduración oligodendroglial. Como resultado (Figura 2), las iOPC se pueden generar en 2 semanas (~95% positivo para el marcador O4) y las iOLs en cuatro semanas (expresando marcadores maduros MBP y PLP1). Curiosamente, encontramos que las iOPC solas secretan una cantidad notable de β amiloide (Aβ), consistente con los datos transcriptómicos independientes que muestran la abundante expresión de la proteína precursora amiloide (APP) y la proteasa de procesamiento β-secretasa (BACE1) en células de linaje oligodendrocitos35,36. Además, nuestro sistema de cocultivo iN-iOPC promueve el envoltorio de axones mediante procesos iOL positivos para MBP y proporciona un apoyo significativo para la formación de sinapsis (Figura 3). Por lo tanto, los protocolos que hemos detallado a continuación tienen ventajas técnicas y biológicas sobre los métodos de cocultivo neurona-oligodendroglia previamente catalogados, y prometen modelar mejor la neurodegeneración en la EA.

<table>
	<tbody>
		<tr>
			<td>Accutase</td>
			<td>STEMCELL Technologies</td>
			<td>7920</td>
			<td></td>
		</tr>
		<tr>
			<td>B27 supplement</td>
			<td>ThermoFisher</td>
			<td>17504044</td>
			<td></td>
		</tr>
		<tr>
			<td>bFGF</td>
			<td>ThermoFisher</td>
			<td>PHG 0266</td>
			<td></td>
		</tr>
		<tr>
			<td>cAMP</td>
			<td>MilliporeSigma</td>
			<td>A9501</td>
			<td></td>
		</tr>
		<tr>
			<td>Clemastine</td>
			<td>MilliporeSigma</td>
			<td>SML0445</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F12 medium</td>
			<td>STEMCELL Technologies</td>
			<td>36254</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>ThermoFisher</td>
			<td>D12345</td>
			<td></td>
		</tr>
		<tr>
			<td>Doxycycline</td>
			<td>MilliporeSigma</td>
			<td>D3072</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>ScienCell</td>
			<td>10</td>
			<td></td>
		</tr>
		<tr>
			<td>H1 human ES cells</td>
			<td>WiCell</td>
			<td>WA01</td>
			<td></td>
		</tr>
		<tr>
			<td>Matrigel</td>
			<td>Corning</td>
			<td>354234</td>
			<td></td>
		</tr>
		<tr>
			<td>mTeSR plus</td>
			<td>STEMCELL Technologies</td>
			<td>5825</td>
			<td></td>
		</tr>
		<tr>
			<td>N2 supplement</td>
			<td>ThermoFisher</td>
			<td>17502001</td>
			<td></td>
		</tr>
		<tr>
			<td>Neurobasal A medium</td>
			<td>ThermoFisher</td>
			<td>10888-022</td>
			<td></td>
		</tr>
		<tr>
			<td>Non Essential Amino Acids</td>
			<td>ThermoFisher</td>
			<td>11140-050</td>
			<td></td>
		</tr>
		<tr>
			<td>PDGF-AA</td>
			<td>R&#38;D Systems</td>
			<td>221-AA-010</td>
			<td></td>
		</tr>
		<tr>
			<td>PEI</td>
			<td>VWR</td>
			<td>71002-812</td>
			<td></td>
		</tr>
		<tr>
			<td>pMDLg/pRRE</td>
			<td>Addgene</td>
			<td>12251</td>
			<td></td>
		</tr>
		<tr>
			<td>Polybrene</td>
			<td>MilliporeSigma</td>
			<td>TR-1003-G</td>
			<td></td>
		</tr>
		<tr>
			<td>pRSV-REV</td>
			<td>Addgene</td>
			<td>12253</td>
			<td></td>
		</tr>
		<tr>
			<td>Puromycin</td>
			<td>ThermoFisher</td>
			<td>A1113803</td>
			<td></td>
		</tr>
		<tr>
			<td>ROCK Inhibitor Y-27632</td>
			<td>STEMCELL Technologies</td>
			<td>72302</td>
			<td></td>
		</tr>
		<tr>
			<td>SAG</td>
			<td>Tocris</td>
			<td>4366</td>
			<td></td>
		</tr>
		<tr>
			<td>STEMdiff Neural Progenitor Freezing Media</td>
			<td>STEMCELL Technologies</td>
			<td>5838</td>
			<td></td>
		</tr>
		<tr>
			<td>STEMdiff SMADi Neural Induction Kit</td>
			<td>STEMCELL Technologies</td>
			<td>8581</td>
			<td></td>
		</tr>
		<tr>
			<td>T3 triiodothyronine</td>
			<td>MilliporeSigma</td>
			<td>T6397</td>
			<td></td>
		</tr>
		<tr>
			<td>Tempo-iOlogo: Human iPSC-derived OPCs</td>
			<td>Tempo BioScience</td>
			<td>SKU102</td>
			<td></td>
		</tr>
		<tr>
			<td>TetO-Ng2-Puro</td>
			<td>Addgene</td>
			<td>52047</td>
			<td></td>
		</tr>
		<tr>
			<td>VSV-G</td>
			<td>Addgene</td>
			<td>12259</td>
			<td></td>
		</tr>
	</tbody>
</table>

generation of human neurons and oligodendrocytes from pluripotent stem cells for modeling neuron-oligodendrocyte interactions

En la enfermedad de Alzheimer (EA) y otros trastornos neurodegenerativos, la insuficiencia oligodendroglial es una característica patológica temprana común, pero la forma en que contribuye al desarrollo y progresión de la enfermedad, particularmente en la materia gris del cerebro, sigue siendo en gran parte desconocida. La disfunción de las células del linaje de oligodendrocitos se caracteriza por deficiencias en la mielinización y alteración de la auto-renovación de las células precursoras de oligodendrocitos (OPC). Estos dos defectos son causados, al menos en parte, por la interrupción de las interacciones entre la neurona y los oligodendrocitos a lo largo de la acumulación de patología. Las OPC dan lugar a oligodendrocitos mielinizantes durante el desarrollo del SNC. En la corteza cerebral madura, las OPC son las principales células proliferativas (que comprenden ~ 5% del total de células cerebrales) y controlan la formación de nueva mielina de una manera dependiente de la actividad neuronal. Tales comunicaciones neuronómicas a oligodendrocitos están significativamente poco estudiadas, especialmente en el contexto de afecciones neurodegenerativas como la EA, debido a la falta de herramientas adecuadas. En los últimos años, nuestro grupo y otros han hecho progresos significativos para mejorar los protocolos actualmente disponibles para generar neuronas funcionales y oligodendrocitos individualmente a partir de células madre pluripotentes humanas. En este manuscrito, describimos nuestros procedimientos optimizados, incluido el establecimiento de un sistema de cocultivo para modelar las conexiones neurona-oligodendrocitos. Nuestros resultados ilustrativos sugieren una contribución inesperada de OPC / oligodendrocitos a la amiloidosis cerebral y la integridad de la sinapsis y destacan la utilidad de esta metodología para la investigación de la EA. Este enfoque reduccionista es una herramienta poderosa para diseccionar las interacciones heterocelulares específicas de la complejidad inherente dentro del cerebro. Se espera que los protocolos que describimos aquí faciliten futuros estudios sobre defectos oligodendrogliales en la patogénesis de la neurodegeneración.

Generación directa de neuronas humanas inducidas a partir de células madre pluripotentes humanas Es muy importante que las células madre pluripotentes humanas iniciales exhiban un alto grado de pluripotencia para la generación exitosa de iNs o iOPCs/iOLs. Por lo tanto, las células deben teñirse para marcadores específicos, como Oct4 y SOX2, antes de iniciar cualquiera de los protocolos de inducción descritos en el presente manuscrito (Figura 1A). Se utilizaron cé...

Watch this Scientific Journal Video about Generation of Human Neurons and Oligodendrocytes from Pluripotent Stem Cells for Modeling Neuron-Oligodendrocyte Interactions at JoVE.com

Generation of Human Neurons and Oligodendrocytes from Pluripotent Stem Cells for Modeling Neuron-Oligodendrocyte Interactions

Las interacciones neurona-glial en la neurodegeneración no se comprenden bien debido a herramientas y métodos inadecuados. Aquí, describimos protocolos optimizados para obtener neuronas inducidas, células precursoras de oligodendrocitos y oligodendrocitos a partir de células madre pluripotentes humanas y proporcionamos ejemplos de los valores de estos métodos para comprender las contribuciones específicas del tipo de célula en la enfermedad de Alzheimer.

Generación de neuronas humanas y oligodendrocitos a partir de células madre pluripotentes para modelar las interacciones neuronal-oligodendrocitos

In Alzheimer&#8217;s disease (AD) and other neurodegenerative disorders, oligodendroglial failure is a common early pathological feature, but how it ...

Research

JoVE Journal

Neuroscience

Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue

Generación de las células troncales nerviosas de desechados Humanos tejido cortical fetal

Neural stem cells (NSCs) reside along the ventricular zone neuroepithelium during the development of the cortical plate. These early progenitors ...

Investigación

Neurociencias

Efficient Derivation of Human Neuronal Progenitors and Neurons from Pluripotent Human Embryonic Stem Cells with Small Molecule Induction

Derivación eficiente de los Derechos Humanos progenitores neuronales y neuronas de pluripotentes células madre embrionarias humanas mediante la inducción de moléculas pequeñas

There is a large unfulfilled need for a clinically-suitable human neuronal cell source for repair or regeneration of the damaged central nervous system ...

The Specification of Telencephalic Glutamatergic Neurons from Human Pluripotent Stem Cells

La especificación de telencefálicas neuronas glutamatérgicas a partir de células madre pluripotentes

Here, a stepwise procedure for efficiently generating telencephalic glutamatergic neurons from human pluripotent stem cells (PSCs) has been described. The ...

A cGMP-applicable Expansion Method for Aggregates of Human Neural Stem and Progenitor Cells Derived From Pluripotent Stem Cells or Fetal Brain Tissue

Un método de expansión de GMPc aplicable para agregados de Neural humano madre y células progenitoras derivadas de células madre pluripotentes o tejido cerebral fetal

A cell expansion technique to amass large numbers of cells from a single specimen for research experiments and clinical trials would greatly benefit the ...

Feeder-free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells

Derivación-Feeder libres de Células Progenitoras de la cresta neural Pluripotentes Células Madre

Human pluripotent stem cells (hPSCs) have great potential for studying human embryonic development, for modeling human diseases in the dish and as a ...

Directed Dopaminergic Neuron Differentiation from Human Pluripotent Stem Cells

Dirigida dopaminérgica Neurona La diferenciación de las células madre pluripotentes humanas

Dopaminergic (DA) neurons in the substantia nigra pars compacta  (also known as A9 DA neurons) are the specific cell type that is lost in ...

Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts

Sencilla generación de una cultura de alto rendimiento de las neuronas inducidas de fibroblastos de piel humana adulta

Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue that is easily ...

Post-differentiation Replating of Human Pluripotent Stem Cell-derived Neurons for High-content Screening of Neuritogenesis and Synapse Maturation

Replacado posterior a la diferenciación de las neuronas derivadas de células madre pluripotentes humanas para la detección de alto contenido de la neuritogénesis y la maduración de la sinapsis

Neurons differentiated in two-dimensional culture from human pluripotent stem-cell-derived neural progenitor cells (NPCs) represent a powerful model ...

Conversion of Human Induced Pluripotent Stem Cells (iPSCs) into Functional Spinal and Cranial Motor Neurons Using PiggyBac Vectors

Conversion of Human Induced Pluripotent Stem Cells iPSCs into Functional Spinal and Cranial Motor Neurons Using PiggyBac Vectors

Conversión de células madre pluripotentes inducidas por humanos (IPSC) en neuronas motoras craneales y espinales funcionales usando vectores PiggyBac

We describe here a method to obtain functional spinal and cranial motor neurons from human induced pluripotent stem cells (iPSCs). Direct conversion into ...

Generation of Oligodendrocytes and Oligodendrocyte-Conditioned Medium for Co-Culture Experiments

Generación de oligodendrocitos y medio condicionado por oligodendrocitos para experimentos de cocultura

In the central nervous system, oligodendrocytes are well-known for their role in axon myelination, that accelerates the propagation of action potentials ...