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

Detta arbete stöddes av bidrag från National Institutes of Health (R00 AG054616 till Y.A.H. och T32 GM136566 till K.C.), Stanford University School of Medicine och ett Siebel Fellowship (tilldelas SC). Y.A.H. är en GFL Translational Professor från Center for Translational Neuroscience vid Brown Institute for Translational Sciences.

1. Mänsklig neuroninduktion från humana pluripotenta stamceller
<ol><li>Lentivirusberedning (~ 5 dagar, detaljerat protokoll som beskrivits tidigare16)<ol><li>Platta ~ 1 miljon HEK293T-celler varje T75-kolv, för att ha dem ~ 40% sammanflytande vid transfektion. Transfektera dem med plasmider som uttrycker tetracyklininducerbar Ngn2 och puromycinresistent gen (PuroR; under samma TetO-promotorkontroll), rtTA och de tre hjälparplasmiderna pRSV-REV, pMDLg/pRRE och VSV-G (12 μg lentiviralt vekto...

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Förutom det fysiska och metaboliska stödet för att stabilisera synapsstrukturerna och för att underlätta den saltatoriska signalledningen genom myelinering, kan oligodendrocytceller forma neuronalt aktivitetsmönster via snabba och dynamiska korssamtal med neuroner 5,6,7. Medan oligodendrogliala svar i AD-patologi ursprungligen betraktades som endast sekundära till inflammation och oxidativa påfrestningar, finns det nu lo...

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

Oligodendrocythärstamningsceller - inklusive oligodendrocytprekursorceller (OPC), myeliniserande oligodendrocyter och övergångstyper däremellan - utgör en stor grupp mänskliga hjärnceller1 som aktivt deltar i många kritiska funktioner för korrekt drift och underhåll av vårt centrala nervsystem under hela neural utveckling och åldrande 2,3,4 . Medan oligodendrocyter är välkända för att producera myelin för att underlätta neuronal aktivitetsöverföring och stödja axonal hälsa i vit substans, är OPC rikliga (~ 5%) i grå substans där myelinering är knapp och utför aktivitetsberoende signalfunktioner för att styra inlärningsbeteende och minnesbildning 5,6,7,8 . Hur oligodendrogliala celler fungerar och dysfunktion i patogenesen av Alzheimers sjukdom (AD) och andra åldersassocierade neurodegenerativa tillstånd har understuderats9. Bristerna i ett lämpligt modellsystem och brister i den allmänna kunskapen för att vägleda en experimentell väg framåt är de främsta orsakerna till denna klyfta.
Mot bakgrund av de senaste genombrotten när det gäller att härleda mänskliga hjärnceller från pluripotenta stamceller, inklusive embryonala stamceller (ES) och inducerade pluripotenta stamceller (iPS), har sådana cellulära modeller i kombination med moderna genredigeringsverktyg framträtt som robusta verktyg för att hantera den invecklade kopplingen mellan cellulära interaktioner i hjärnan och kan visa människospecifika sjukdomsmanifestationer10, 11. Med tanke på att enskilda hjärncellstyper kan uppvisa distinkta och till och med motstridiga effekter inför samma AD-främjande förhållanden12,13, erbjuder denna stamcellsmetodik unikt celltypspecifik information som tidigare har missats med hjälp av etablerade in vivo- eller in vitro-modeller som endast ger aggregerade avläsningar från samlingar av hjärncellstyper. Under det senaste decenniet har ett stort antal tillförlitliga protokoll utvecklats för att generera mänskliga neuroner från transdifferentiering av ES / iPS-celler eller direkt omvandling från andra terminalt differentierade celltyper (t.ex. fibroblaster)14,15. I synnerhet kan tillämpningen av viktiga neurogena transkriptionsfaktorer (t.ex. neurogenin 2, Ngn2)16 på humana pluripotenta stamceller generera en homogen population av väl karakteriserade neuronala celltyper för rena kulturer utan behov av coculturing med gliaceller12,17,18. För inducerade humana oligodendrocyter finns det några publicerade protokoll som kan generera funktionella celler som mycket liknar deras primära motsvarigheter, med ett brett spektrum av effektivitet och efterfrågan i tid och resurser 19,20,21,22,23,24,25,26,27,28 . Hittills har inget av dessa protokoll tillämpats för att undersöka hur oligodendrogliala celler svarar på och påverkar AD-patogenes.
Här beskriver vi våra förbättrade protokoll för enstaka och blandade kulturer av humaninducerade neuroner (iNs) och OPCs/oligodendrocyter (iOPCs/iOLs). iN-protokollet som beskrivs här är baserat på den allmänt använda Ngn2-metoden16 och har den ytterligare funktionen att vara gliafri. De resulterande iN är homogena och liknar starkt de kortikala skiktet 2/3 excitatoriska neuroner, med karakteristisk pyramidal morfologi, genuttrycksmönster och elektrofysiologiska egenskaper17,18 (Figur 1). För att övervinna några av de grundläggande hindren för riktad differentiering av pluripotenta stamceller har vi utvecklat en enkel och effektiv metod för lågdos dimetylsulfoxid (DMSO) förbehandling29,30 och rapporterat en ökad benägenhet hos humana ES / iPS-celler att transdifferentiera till iOPCs och iOLs31, baserat på ett allmänt anpassat protokoll av Douvaras och Fossati 32 . Vi har ytterligare förenklat protokollet och införlivat en robust differentieringsfrämjande förening, clemastine 7,33,34, för att påskynda processen för oligodendroglial mognad. Som ett resultat (figur 2) kan iOPC: erna genereras på 2 veckor (~ 95% positiva för markören O4) och iOL på fyra veckor (uttrycker mogna markörer MBP och PLP1). Intressant nog fann vi att iOPCs ensamma utsöndrar en anmärkningsvärd mängd amyloid-β (Aβ), i överensstämmelse med de oberoende transkriptomiska data som visar det rikliga uttrycket av amyloidprekursorproteinet (APP) och bearbetningsproteas β-sekretas (BACE1) i oligodendrocythärstamningsceller35,36. Dessutom främjar vårt iN-iOPC-samodlingssystem inneslutningen av axoner genom MBP-positiva iOL-processer och ger betydande stöd för synapsbildning (figur 3). Således har de protokoll som vi har beskrivit nedan tekniska och biologiska fördelar jämfört med tidigare katalogiserade neuron-oligodendroglia co-culturing-metoder och har ett löfte om bättre modellering av neurodegenerationen i AD.

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			<td>Fetal Bovine Serum</td>
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			<td>ThermoFisher</td>
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			<td>Non Essential Amino Acids</td>
			<td>ThermoFisher</td>
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			<td>STEMdiff Neural Progenitor Freezing Media</td>
			<td>STEMCELL Technologies</td>
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			<td>STEMdiff SMADi Neural Induction Kit</td>
			<td>STEMCELL Technologies</td>
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			<td>MilliporeSigma</td>
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			<td>Tempo-iOlogo: Human iPSC-derived OPCs</td>
			<td>Tempo BioScience</td>
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generation of human neurons and oligodendrocytes from pluripotent stem cells for modeling neuron-oligodendrocyte interactions

Vid Alzheimers sjukdom (AD) och andra neurodegenerativa sjukdomar är oligodendroglial misslyckande ett vanligt tidigt patologiskt drag, men hur det bidrar till sjukdomsutveckling och progression, särskilt i hjärnans grå substans, är fortfarande i stort sett okänt. Dysfunktionen hos oligodendrocytceller kännetecknas av brister i myelinering och nedsatt självförnyelse av oligodendrocytprekursorceller (OPC). Dessa två defekter orsakas åtminstone delvis av störningen av interaktioner mellan neuron och oligodendrocyter längs uppbyggnaden av patologi. OPC ger upphov till myeliniserande oligodendrocyter under CNS-utveckling. I den mogna hjärnbarken är OPC de viktigaste proliferativa cellerna (som omfattar ~ 5% av de totala hjärncellerna) och kontrollerar ny myelinbildning på ett neuralt aktivitetsberoende sätt. Sådan neuron-till-oligodendrocytkommunikation är signifikant understuderad, särskilt i samband med neurodegenerativa tillstånd såsom AD, på grund av bristen på lämpliga verktyg. Under de senaste åren har vår grupp och andra gjort betydande framsteg för att förbättra för närvarande tillgängliga protokoll för att generera funktionella neuroner och oligodendrocyter individuellt från humana pluripotenta stamceller. I detta manuskript beskriver vi våra optimerade procedurer, inklusive upprättandet av ett samodlingssystem för att modellera neuron-oligodendrocytanslutningarna. Våra illustrativa resultat tyder på ett oväntat bidrag från OPCs / oligodendrocyter till hjärnans amyloidos och synapsintegritet och belyser nyttan av denna metod för AD-forskning. Detta reduktionistiska tillvägagångssätt är ett kraftfullt verktyg för att dissekera de specifika heterocellulära interaktionerna ur den inneboende komplexiteten i hjärnan. De protokoll vi beskriver här förväntas underlätta framtida studier av oligodendrogliala defekter i patogenesen av neurodegeneration.

Direkt generering av humaninducerade nervceller från humana pluripotenta stamceller Det är mycket viktigt att de ursprungliga humana pluripotenta stamcellerna uppvisar en hög grad av pluripotens för framgångsrik generering av iNs eller iOPCs/iOLs. Därför bör celler färgas för specifika markörer, såsom okt4 och SOX2, innan någon av de induktionsprotokoll som beskrivs i detta manuskript (figur 1A) påbörjas. Mänskliga H1-celler användes för att erhålla ind...

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

Neuron-glial interaktioner i neurodegeneration är inte väl förstådda på grund av otillräckliga verktyg och metoder. Här beskriver vi optimerade protokoll för att erhålla inducerade neuroner, oligodendrocytprekursorceller och oligodendrocyter från humana pluripotenta stamceller och ger exempel på värdena för dessa metoder för att förstå celltypspecifika bidrag vid Alzheimers sjukdom.

Generering av humana neuroner och oligodendrocyter från pluripotenta stamceller för modellering av neuron-oligodendrocytinteraktioner

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

The protocols described here, are expected to facilitate future studies on oligodendroglial defects, in the pathogenesis of neuro degeneration. This is an improved method that can be used to generate functional human oligodendrocyte lineage cells in only three to four weeks, and establish a co-culture system to model the neuron oligodendrocyte connections. The protocol can be used to study not only Alzheimer's disease, but any other disease associated aspect of neuron oligodendrocyte interactions.To begin, plate approximately one million HEK293T cells in each T75 flask to achieve 40%confluency, then transfect them with plasmids expressing tetracycline inducible NGN2 and puromycin resistant tetracycline transactivator gene, along with the three helper plasmids. Use PEI for transfection. Change the media after 16 hours.Harvest released viral particles by collecting culture media every day and replacing it with fresh media for three days. Pool the collected media for purification, then filter the virus through a 0.22 micrometer filter and centrifuge at 49, 000 times G for 90 minutes. After centrifugation, re-suspend the pellet in the appropriate volume of PBS glucose.Use commercially available H1 human ES cells to perform neuron induction. Culture the cells on extracellular matrix solution coated 6-well plates with ESL maintenance medium and incubate the plates at 37 degrees Celsius with 5%carbon dioxide. Two days before induction of differentiation, detach ES cells by adding one milliliter of cell detachment solution and incubating at room temperature for 10 minutes.Transfer the cells to a tube, then wash the well with two milliliters of media and add it to the same tube. Centrifuge the cells at 300 times G for five minutes. Then re-suspend the pellet in media and plate the cells onto matrix coated 6-well plates at the seating density of one times 10 to the fifth cells per well.On the next day, add lentiviruses, expressing NGN2 plus puro. R, and tetracycline transactivator together with polybrene in fresh ES cell maintenance medium. On day zero, add two micrograms per milliliter doxycycline in DMEM/F-12 medium supplemented with N2 and without morphogens.On day one, add puromycin in fresh DMEM/F-12, plus N2 and doxycycline to the final concentration of one microgram per milliliter. Select the transduced cells in puromycin for at least 24 hours. On day two, detach differentiating neurons with cell detachment solution and re-plate them on 24-well plates coated with matrix solution.Maintain them in MBA B27 medium without doxycycline. Culture pure induced neurons on the plates, coated with extracellular matrix-based solutions. To perform neural progenitor cell generation, culture H1 human ES cells and transdifferentiate them into neural progenitor cells, as described in the text manuscript.On day negative one, seed 0.5 to one million cells in ES cell maintenance medium into each well of a 6-well plate coated with growth factor reduced matrix solution. On day zero, treat cells for 24 hours with ES cell maintenance medium supplemented with 2%DMSO. On days one through six, change the full media with warm neural induction medium containing some add inhibitors from a commercial kit.On day seven, passage NPCs using cell detachment solution and plate them at a seating density of one to two times 10 to the fifth cells per well of a 24-well plate. Assay the differentiation efficiency by immunohistochemical staining for absence of pluripotentcy marker and presence of NPC markers. To generate the oligodendrocyte precursor cells, plate the NPCs in warm neural induction medium with some add inhibitors from a commercial kit.On day eight, prepare a solution of 1%DMSO in the OPC differentiation medium, and treat the plated NPCs for 24 hours. On day nine, replace the media with fresh OPC differentiation medium without DMSO, then feed the cells every other day until day 15. If the the cells reach confluence before day 15, passage them to the seating density of one to two times 10 to the fifth cells per well.On day 14, plate OPCs in OPC differentiation medium at a density of one to two times 10 to the fifth cells per well, in a 24-well plate. Test cells for the presence of OPC specific markers by IHC staining or qPCR, and for the absence of NPC markers. On day 15, replace media with OL maturation medium.Change the medium every other day or every day. When cells reach 90%confluence, split them at a one to three ratio for up to two passages or until cell division slows down substantially. If OPCs divide too fast and reach confluencey in less than three days, add Ara-C at a concentration of two to five micromolar for one to three days.On day 14, plate the IOPCs at a density of one times 10 to the fifth cells per well in a 24-well plate in OPC differentiation medium. On the next day, detach the induced human neurons with cell detachment solution. Add neurons onto the cultured OPCs, plating at the seating density of two times 10 to the fifth cells per well.Use co-culture medium containing neurobasal A medium, 2%B27 supplement, and 100 nanograms per milliliter T3 triiodothyronine. Change the medium on the next day and then every other day. If OPCs proliferate too fast and reach confluency in less than three days, add two to five micromolar Ara-C.It is important that the starting human pluripotent stem cells exhibit a high degree of pluripotentcy for the successful generation of induced neurons, or OPCs and oligodendrocytes. Therefore, cells should be stained for specific markers, such as OCT-4 and SOX2, before starting either of the induction protocols. Human H1 cells were used to obtain induced excitatory forebrain neurons.Pure neurons in culture started expressing synapse in one at day 14 to 16. IOPCs were generated in two weeks and mature oligodendrocytes in four to five weeks. DMSO treatment enriched the number of cells in the early G1 phase for better signaling Integration.Specific OPC markers were detected as early as two weeks after plating of ES H1 cells. The IOPC population at two weeks was fairly homogenous, with over 95%of cells positive for 04 staining and a high level of immunoreactivity for other markers. After initiation of oligodendrocyte maturation, specific oligodendrocyte markers were detected starting at day 28.The ideal plating densities for induced neurons, and IOPCs, need to be determined with a series of cell number titrations to achieve proper differentiation. After four weeks, in co-cultures, the IOPCs should be adequately differentiated into oligodendrocytes that are positive for specific markers such as MBP. This co-culture system can robustly boost the number of synapses, indicating that the IOPCs provide a neuronal support through physical contact or release of trophic factors.This described procedure can be further sophisticated by adding different brain cell types. For example, stem cells derived human astrocytes to model novel aspects of neuron glial interactions in health and disease.

Research

JoVE Journal

Neuroscience

Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue

Generering av neuronala stamceller från kasserade mänskliga foster Bark-Tissue

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

Effektiv Derivering av mänskliga neurala stamceller och nervceller från Pluripotenta mänskliga embryonala stamcellslinjer med liten molekyl Induktion

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

Specifikationen av Telencephalic glutamaterg nervceller från mänskliga pluripotenta stamceller

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

En cGMP-tillämplig Expansion Metod för aggregat av mänskliga neurala Stem och stamceller härledas från pluripotenta stamceller eller fosterhjärnvävnad

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

Feeder fria Härledning av neurallisten progenitorceller från Human pluripotenta stamceller

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

Riktad dopaminerga nerv Differentiering från Human pluripotenta stamceller

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

Enkla Generation av en hög avkastning kultur av inducerade nervceller från mänsklig vuxen hud fibroblaster

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 ...

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

Omvandling av mänskliga inducerade pluripotenta stamceller (iPSCs) till funktionella spinal och kranial motor neuron använda PiggyBac vektorer

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

Derivation, Expansion, Cryopreservation and Characterization of Brain Microvascular Endothelial Cells from Human Induced Pluripotent Stem Cells

Härledning, expansion, kryopreservation och karakterisering av hjärnans mikrovaskulära endotelceller från mänskliga inducerade pluripotenta stamceller

Brain microvascular endothelial cells (BMECs) can be differentiated from human induced pluripotent stem cells (iPSCs) to develop ex vivo cellular models ...

Establishment of an Electrophysiological Platform for Modeling ALS with Regionally-Specific Human Pluripotent Stem Cell-Derived Astrocytes and Neurons

Etablering av en elektrofysiologisk plattform för modellering av ALS med regionalt specifika humana pluripotenta stamcellshärledda astrocyter och neuroner

Human pluripotent stem cell-derived astrocytes (hiPSC-A) and neurons (hiPSC-N) provide a powerful tool for modeling Amyotrophic Lateral Sclerosis (ALS) ...