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

Dit werk werd ondersteund door de subsidies van National Institutes of Health (R00 AG054616 aan Y.A.H. en T32 GM136566 aan K.C.), Stanford University School of Medicine en een Siebel Fellowship (toegekend aan S.C.). Y.A.H. is een GFL Translational Professor van het Center for Translational Neuroscience in het Brown Institute for Translational Sciences.

1. Inductie van menselijke neuronen uit menselijke pluripotente stamcellen
<ol><li>Lentivirus voorbereiding (~ 5 dagen, gedetailleerd protocol zoals eerder beschreven16)<ol><li>Plaat ~ 1 miljoen HEK293T-cellen per T75-kolf, om ze ~ 40% confluent te hebben bij het uitvoeren van transfectie. Transfecteer ze met plasmiden die tetracycline-induceerbare Ngn2 en puromycine-resistent gen (PuroR; onder dezelfde TetO-promotorcontrole) tot expressie brengen, rtTA en de drie helperplasmiden pRSV-REV, pMDLg...

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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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Naast de fysieke en metabole ondersteuning om de synapsstructuren te stabiliseren en de zoute signaalgeleiding door myelinisatie te vergemakkelijken, kunnen oligodendrocytenafstammingscellen een neuronaal activiteitspatroon vormen via snelle en dynamische kruisgesprekken met neuronen 5,6,7. Terwijl in de AD-pathologie de oligodendrogliale responsen aanvankelijk werden beschouwd als slechts secundair aan ontsteking en oxidatieve ...

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

Oligodendrocytenafstammingscellen - inclusief oligodendrocytenvoorlopercellen (OPC's), myeliniserende oligodendrocyten en overgangstypen daartussenin - vormen een belangrijke groep menselijke hersencellen1 die actief deelnemen aan vele kritieke functies voor de juiste werking en het onderhoud van ons centrale zenuwstelsel tijdens de neurale ontwikkeling en veroudering 2,3,4 . Hoewel oligodendrocyten bekend staan om het produceren van myeline om de overdracht van neuronale activiteit te vergemakkelijken en de axonale gezondheid in witte stof te ondersteunen, zijn OPC's overvloedig (~ 5%) in grijze stof waar myelinisatie schaars is en voeren activiteitsafhankelijke signaleringsfuncties uit om leergedrag en geheugenvorming te regelen 5,6,7,8 . Hoe oligodendrogliale cellen functioneren en disfunctie in de pathogenese van de ziekte van Alzheimer (AD) en andere leeftijdsgebonden neurodegeneratieve aandoeningen is onderbelicht9. De tekortkomingen van een geschikt modelsysteem en tekortkomingen in de algemene kennis om een experimenteel pad voorwaarts te begeleiden, zijn de belangrijkste redenen voor deze kloof.
In het licht van de laatste doorbraken in het afleiden van menselijke hersencellen uit pluripotente stamcellen, waaronder embryonale stamcellen (ES) en geïnduceerde pluripotente stamcellen (iPS), zijn dergelijke cellulaire modellen in combinatie met moderne genbewerkingstools naar voren gekomen als robuuste hulpmiddelen om de ingewikkelde nexus van cellulaire interacties in de hersenen aan te pakken, en zijn ze in staat om mensspecifieke ziekteverschijnselen aan te tonen10, 11. Gezien het feit dat individuele hersenceltypen verschillende en zelfs conflicterende effecten kunnen vertonen in het licht van dezelfde AD-bevorderende aandoeningen12,13, biedt deze stamcelmethodologie op unieke wijze celtypespecifieke informatie die eerder is gemist met behulp van gevestigde in vivo of in vitro modellen die alleen geaggregeerde uitlezingen bieden van verzamelingen hersenceltypen. In het afgelopen decennium zijn een groot aantal betrouwbare protocollen ontwikkeld om menselijke neuronen te genereren uit transdifferentiatie van ES / iPS-cellen of directe conversie van andere terminaal gedifferentieerde celtypen (bijv. Fibroblasten)14,15. In het bijzonder kan de toepassing van belangrijke neurogene transcriptiefactoren (bijv. neurogenine 2, Ngn2)16 op menselijke pluripotente stamcellen een homogene populatie van goed gekarakteriseerde neuronale celtypen genereren voor zuivere culturen zonder dat er behoefte is aan coculturing met gliacellen 12,17,18. Voor geïnduceerde menselijke oligodendrocyten zijn er een paar gepubliceerde protocollen die functionele cellen kunnen genereren die sterk lijken op hun primaire tegenhangers, met een breed scala aan efficiëntie en vraag in tijd en middelen 19,20,21,22,23,24,25,26,27,28 . Tot op heden is geen van deze protocollen toegepast om te onderzoeken hoe oligodendrogliale cellen reageren op en invloed hebben op AD-pathogenese.
Hier beschrijven we onze verbeterde protocollen voor enkelvoudige en gemengde culturen van door de mens geïnduceerde neuronen (iN's) en OPC's/oligodendrocyten (iOPCs/iOL's). Het hier beschreven iN-protocol is gebaseerd op de veelgebruikte Ngn2-benadering16 en heeft als extra eigenschap gliavrij te zijn. De resulterende iNs zijn homogeen en lijken sterk op de corticale laag 2/3 exciterende neuronen, met karakteristieke piramidale morfologie, genexpressiepatroon en elektrofysiologische kenmerken17,18 (figuur 1). Om enkele van de fundamentele barrières bij gerichte differentiatie van pluripotente stamcellen te overwinnen, hebben we een eenvoudige en effectieve methode ontwikkeld voor de voorbehandeling van lage doses dimethylsulfoxide (DMSO)29,30, en rapporteerden we een verhoogde neiging van menselijke ES / iPS-cellen om te transdifferentiëren in iOPCs en iOL's31, gebaseerd op een breed aangepast protocol van Douvaras en Fossati32 . We hebben het protocol verder vereenvoudigd en een robuuste differentiatiebevorderende verbinding, clemastine 7,33,34, opgenomen om het proces van oligodendrogliale rijping te versnellen. Als gevolg hiervan (figuur 2) kunnen de iOPCs worden gegenereerd in 2 weken (~ 95% positief voor de marker O4) en iOL's in vier weken (met volwassen markers MBP en PLP1). Interessant is dat we ontdekten dat iOPCs alleen een opmerkelijke hoeveelheid amyloïde-β (Aβ) afscheiden, consistent met de onafhankelijke transcriptomische gegevens die de overvloedige expressie van het amyloïde precursoreiwit (APP) en het verwerkingsprotease β-secretase (BACE1) in oligodendrocytenlijncellen35,36 aantonen. Bovendien bevordert ons iN-iOPC cocultuursysteem de opwarming van axonen door MBP-positieve iOL-processen en biedt het aanzienlijke ondersteuning voor synapsvorming (figuur 3). De protocollen die we hieronder hebben beschreven, hebben dus technische en biologische voordelen ten opzichte van eerder gecatalogiseerde neuron-oligodendroglia co-cultiveringsmethoden en houden een belofte in bij het beter modelleren van de neurodegeneratie in AD.

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			<td>STEMCELL Technologies</td>
			<td>7920</td>
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			<td>ThermoFisher</td>
			<td>17504044</td>
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			<td>bFGF</td>
			<td>ThermoFisher</td>
			<td>PHG 0266</td>
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			<td>MilliporeSigma</td>
			<td>A9501</td>
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		<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>
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			<td>DMSO</td>
			<td>ThermoFisher</td>
			<td>D12345</td>
			<td></td>
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			<td>Doxycycline</td>
			<td>MilliporeSigma</td>
			<td>D3072</td>
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			<td>Fetal Bovine Serum</td>
			<td>ScienCell</td>
			<td>10</td>
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			<td>H1 human ES cells</td>
			<td>WiCell</td>
			<td>WA01</td>
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			<td>Matrigel</td>
			<td>Corning</td>
			<td>354234</td>
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			<td>mTeSR plus</td>
			<td>STEMCELL Technologies</td>
			<td>5825</td>
			<td></td>
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			<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>
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			<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>
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			<td>pRSV-REV</td>
			<td>Addgene</td>
			<td>12253</td>
			<td></td>
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			<td>Puromycin</td>
			<td>ThermoFisher</td>
			<td>A1113803</td>
			<td></td>
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		<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

Bij de ziekte van Alzheimer (AD) en andere neurodegeneratieve aandoeningen is oligodendrogliaal falen een veel voorkomend vroeg pathologisch kenmerk, maar hoe het bijdraagt aan de ontwikkeling en progressie van de ziekte, met name in de grijze stof van de hersenen, blijft grotendeels onbekend. De disfunctie van oligodendrocytenafstammingscellen wordt gekenmerkt door tekortkomingen in myelinisatie en verminderde zelfvernieuwing van oligodendrocytenvoorlopercellen (OPC's). Deze twee defecten worden ten minste gedeeltelijk veroorzaakt door de verstoring van interacties tussen neuronen en oligodendrocyten langs de opbouw van pathologie. OPC's geven aanleiding tot myeliniserende oligodendrocyten tijdens de ontwikkeling van het CZS. In de volwassen hersenschors zijn OPC's de belangrijkste proliferatieve cellen (bestaande uit ~ 5% van de totale hersencellen) en controleren nieuwe myelinevorming op een neurale activiteitsafhankelijke manier. Dergelijke neuron-naar-oligodendrocytencommunicatie wordt aanzienlijk onderbelicht, vooral in de context van neurodegeneratieve aandoeningen zoals AD, vanwege het gebrek aan geschikte hulpmiddelen. In de afgelopen jaren hebben onze groep en anderen aanzienlijke vooruitgang geboekt bij het verbeteren van de momenteel beschikbare protocollen om functionele neuronen en oligodendrocyten individueel te genereren uit menselijke pluripotente stamcellen. In dit manuscript beschrijven we onze geoptimaliseerde procedures, waaronder het opzetten van een co-kweeksysteem om de neuron-oligodendrocytenverbindingen te modelleren. Onze illustratieve resultaten suggereren een onverwachte bijdrage van OPC's / oligodendrocyten aan de amyloïdose van de hersenen en synapsintegriteit en benadrukken het nut van deze methodologie voor AD-onderzoek. Deze reductionistische benadering is een krachtig hulpmiddel om de specifieke hetero-cellulaire interacties te ontleden uit de inherente complexiteit in de hersenen. De protocollen die we hier beschrijven, zullen naar verwachting toekomstige studies naar oligodendrogliale defecten in de pathogenese van neurodegeneratie vergemakkelijken.

Directe generatie van door de mens geïnduceerde neuronen uit menselijke pluripotente stamcellen Het is van groot belang dat de startende menselijke pluripotente stamcellen een hoge mate van pluripotentie vertonen voor een succesvolle generatie van iN's of iOPCs/iOL's. Daarom moeten cellen worden gekleurd voor specifieke markers, zoals Oct4 en SOX2, voordat ze beginnen met een van de inductieprotocollen die in dit manuscript worden beschreven (figuur 1A). Menselijke H1-ce...

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

De neuron-gliale interacties bij neurodegeneratie worden niet goed begrepen als gevolg van ontoereikende hulpmiddelen en methoden. Hier beschrijven we geoptimaliseerde protocollen om geïnduceerde neuronen, oligodendrocytenvoorlopercellen en oligodendrocyten te verkrijgen uit menselijke pluripotente stamcellen en geven we voorbeelden van de waarden van deze methoden bij het begrijpen van celtypespecifieke bijdragen bij de ziekte van Alzheimer.

Generatie van menselijke neuronen en oligodendrocyten uit pluripotente stamcellen voor het modelleren van neuron-oligodendrocyteninteracties

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

Generatie van neurale stamcellen uit afgedankte menselijke foetale Cortical weefsel

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

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

Efficiënte Afleiding van de Mens Neuronale voorouders en neuronen van pluripotente menselijke embryonale stamcellen met op kleine moleculen Inductie

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

De Specificatie van Telencephalic glutamaat neuronen van menselijke pluripotente stamcellen

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

Een cGMP-toepassing Uitbreiding Methode voor Aggregaten van menselijke neurale stamcellen en stamcellen afgeleid uit pluripotente stamcellen of foetaal hersenweefsel

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

Voedingsvrije Afleiding van neurale stamcellen uit humane pluripotente stamcellen

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

Geregisseerd Dopaminerge Neuron Differentiatie van menselijke pluripotente stamcellen

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

Eenvoudige generatie van de cultuur van een hoog rendement van geïnduceerde neuronen van menselijke volwassen huid fibroblasten

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

Na differentiatie Herbeplating van menselijke pluripotente stamcel afkomstige neuronen voor het screenen van hoge inhoud van Neuritogenese en Synapse rijping

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

Conversie van menselijke geïnduceerde pluripotente stamcellen (iPSCs) in functionele spinale en craniale motor neuronen met behulp van PiggyBac vectoren

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

Generatie oligodendrocyten en oligodendrocyte-geconditioneerde medium voor co-cultuurexperimenten

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