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

Bu çalışma, Ulusal Sağlık Enstitüleri (R00 AG054616'dan Y.A.H.'ye ve T32 GM136566'dan K.C.'ye), Stanford Üniversitesi Tıp Fakültesi ve Siebel Bursu'ndan (S.C.'ye verildi) gelen hibelerle desteklenmiştir. Y.A.H., Brown Translasyonel Bilimler Enstitüsü'ndeki Translasyonel Sinirbilim Merkezi'nden GFL Translasyonel Profesörüdür.

1. İnsan pluripotent kök hücrelerinden insan nöron indüksiyonu
<ol><li>Lentivirüs hazırlığı (~ 5 gün, daha önce açıklandığı gibi ayrıntılı protokol16)<ol><li>Her T75 şişesinde ~1 milyon HEK293T hücresi plakası, transfeksiyon yaparken ~% 40 oranında bir araya gelmelerini sağlar. Tetrasiklin ile indüklenebilir Ngn2 ve püromisine dirençli geni (PuroR; aynı TetO promotör kontrolü altında), rtTA ve üç yardımcı plazmid pRSV-REV, pMDLg / pRRE ve VSV-G'yi (12 μg len...

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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. C., 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=Defined+and+scalable+differentiation+of+human+oligodendrocyte+precursors+from+pluripotent+stem+cells+in+a+3D+culture+system.">Defined and scalable differentiation of human oligodendrocyte precursors from pluripotent stem cells in a 3D culture system.</a> Stem Cell Reports. 8 (6), 1770-1783 (2017).</li><li>Hu, B. Y., Du, Z. W., Li, X. J., Ayala, M., Zhang, S. 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Sinaps yapılarını stabilize etmek ve miyelinasyon yoluyla tuzlayıcı sinyal iletimini kolaylaştırmak için fiziksel ve metabolik desteğe ek olarak, oligodendrosit soy hücreleri, nöronlarla hızlı ve dinamik çapraz görüşmeler yoluyla nöronal aktivite paternini şekillendirebilir 5,6,7. AD patolojisinde oligodendroglial yanıtlar başlangıçta inflamasyon ve oksidatif streslere ikincil olarak kabul edilirken, şimd...

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

Oligodendrosit soy hücreleri - oligodendrosit öncü hücreleri (OPC'ler), miyelinizan oligodendrositler ve aralarındaki geçiş tipleri dahil - nöral gelişim ve yaşlanma boyunca merkezi sinir sistemimizin düzgün çalışması ve bakımı için birçok kritik fonksiyona aktif olarak katılan önemli bir insan beyin hücresi grubunu oluşturur1 2,3,4 . Oligodendrositler, nöronal aktivite iletimini kolaylaştırmak ve beyaz cevherde aksonal sağlığı desteklemek için miyelin üretmesiyle iyi bilinirken, OPC'ler, miyelinasyonun az olduğu gri maddede bol miktarda (~% 5) bulunur ve öğrenme davranışını ve hafıza oluşumunu yönetmek için aktiviteye bağlı sinyal işlevlerini yerine getirir 5,6,7,8 . Alzheimer hastalığı (AD) ve diğer yaşa bağlı nörodejeneratif durumların patogenezinde oligodendroglial hücrelerin nasıl işlev gördüğü ve işlev bozukluğu az çalışılmıştır9. Uygun bir model sistemin yetersizlikleri ve deneysel bir yolu ileriye götürecek genel bilgi eksiklikleri bu boşluğun başlıca nedenleridir.
İnsan beyin hücrelerinin, embriyonik kök (ES) ve indüklenmiş pluripotent kök (iPS) hücreler de dahil olmak üzere pluripotent kök hücrelerden türetilmesindeki en son atılımların ışığında, modern gen düzenleme araçlarıyla birlikte bu tür hücresel modeller, beyindeki hücresel etkileşimlerin karmaşık bağlantı noktasını ele almak için sağlam araçlar olarak ortaya çıkmıştır ve insana özgü hastalık belirtilerini gösterebilmektedir10, 11. Bireysel beyin hücresi tiplerinin, aynı AD'yi teşvik eden koşullar12,13 karşısında farklı ve hatta çelişkili etkiler gösterebileceği göz önüne alındığında, bu kök hücre metodolojisi, daha önce beyin hücresi tiplerinin koleksiyonlarından yalnızca toplu okumalar sağlayan in vivo veya in vitro modeller kullanılarak gözden kaçırılmış olan hücre tipine özgü bilgileri benzersiz bir şekilde sunar. Son on yılda, ES / iPS hücrelerinin trans-farklılaşmasından veya diğer ölümcül olarak farklılaşmış hücre tiplerinden (örneğin, fibroblastlar) doğrudan dönüşümden insan nöronları üretmek için çok sayıda güvenilir protokol geliştirilmiştir 14,15. Özellikle, anahtar nörojenik transkripsiyon faktörlerinin (örneğin, nörogenin 2, Ngn2)16'nın insan pluripotent kök hücrelerine uygulanması, glial hücrelerle pıhtılaşmaya gerek kalmadan saf kültürler için iyi karakterize edilmiş nöronal hücre tiplerinin homojen bir popülasyonunu oluşturabilir12,17,18. İndüklenmiş insan oligodendrositleri için, birincil meslektaşlarına çok benzeyen fonksiyonel hücreler üretebilen, zaman ve kaynaklarda geniş bir verimlilik ve talep yelpazesine sahip birkaç yayınlanmış protokol vardır 19,20,21,22,23,24,25,26,27,28 . Bugüne kadar, oligodendroglial hücrelerin AD patogenezine nasıl tepki verdiğini ve onu nasıl etkilediğini araştırmak için bu protokollerin hiçbiri uygulanmamıştır.
Burada, insan kaynaklı nöronların (iN'ler) ve OPC'lerin / oligodendrositlerin (iOPC'ler / iOL'ler) tek ve karışık kültürleri için geliştirilmiş protokollerimizi açıklıyoruz. Burada açıklanan iN protokolü, yaygın olarak kullanılan Ngn2 yaklaşımı16'ya dayanmaktadır ve glia-free olma özelliğine sahiptir. Ortaya çıkan iN'ler homojendir ve karakteristik piramidal morfoloji, gen ekspresyon paterni ve elektrofizyolojik özellikler17,18 ile kortikal tabaka 2/3 uyarıcı nöronlara oldukça benzemektedir (Şekil 1). Pluripotent kök hücrelerin yönlendirilmiş farklılaşmasındaki bazı temel engellerin üstesinden gelmek için, düşük doz dimetil sülfoksit (DMSO) ön tedavisi29,30 için basit ve etkili bir yöntem geliştirdik ve Douvaras ve Fossati32 tarafından geniş çapta uyarlanmış bir protokole dayanarak, insan ES / iPS hücrelerinin iOPC'lere ve iOL'lere31'e dönüşme eğiliminin arttığını bildirdik. . Protokolü daha da basitleştirdik ve oligodendroglial olgunlaşma sürecini hızlandırmak için sağlam bir farklılaşmayı teşvik eden bileşik olan clemastine 7,33,34'ü dahil ettik. Sonuç olarak (Şekil 2), iOPC'ler 2 hafta içinde (O4 belirteci için ~% 95 pozitif) ve iOL'ler dört hafta içinde (olgun belirteçler MBP ve PLP1'i ifade eder) üretilebilir. İlginç bir şekilde, iOPC'lerin tek başına kayda değer miktarda amiloid-β (Aβ) salgıladığını, amiloid öncü proteininin (APP) bol miktarda ekspresyonunu ve oligodendrositsoy hücrelerinde proteaz β-sekretazın (BACE1) işlendiğini gösteren bağımsız transkriptomik verilerle tutarlı olduğunu bulduk. Ayrıca, iN-iOPC ko-kültür sistemimiz, aksonların MBP-pozitif iOL süreçleri ile kaplanmasını teşvik eder ve sinaps oluşumu için önemli destek sağlar (Şekil 3). Bu nedenle, aşağıda detaylandırdığımız protokoller, daha önce kataloglanmış nöron-oligodendroglia ortak kültürleme yöntemlerine göre teknik ve biyolojik avantajlara sahiptir ve AD'de nörodejenerasyonun daha iyi modellenmesinde bir umut vaat etmektedir.

<table>
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			<td>7920</td>
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			<td>Clemastine</td>
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			<td>DMEM/F12 medium</td>
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			<td>Fetal Bovine Serum</td>
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			<td>H1 human ES cells</td>
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			<td>WA01</td>
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			<td>5825</td>
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			<td>N2 supplement</td>
			<td>ThermoFisher</td>
			<td>17502001</td>
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			<td>Neurobasal A medium</td>
			<td>ThermoFisher</td>
			<td>10888-022</td>
			<td></td>
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			<td>Non Essential Amino Acids</td>
			<td>ThermoFisher</td>
			<td>11140-050</td>
			<td></td>
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			<td>PDGF-AA</td>
			<td>R&#38;D Systems</td>
			<td>221-AA-010</td>
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			<td>PEI</td>
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			<td>12251</td>
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			<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>
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			<td>SAG</td>
			<td>Tocris</td>
			<td>4366</td>
			<td></td>
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		<tr>
			<td>STEMdiff Neural Progenitor Freezing Media</td>
			<td>STEMCELL Technologies</td>
			<td>5838</td>
			<td></td>
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		<tr>
			<td>STEMdiff SMADi Neural Induction Kit</td>
			<td>STEMCELL Technologies</td>
			<td>8581</td>
			<td></td>
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		<tr>
			<td>T3 triiodothyronine</td>
			<td>MilliporeSigma</td>
			<td>T6397</td>
			<td></td>
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		<tr>
			<td>Tempo-iOlogo: Human iPSC-derived OPCs</td>
			<td>Tempo BioScience</td>
			<td>SKU102</td>
			<td></td>
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		<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>
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	</tbody>
</table>

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

Alzheimer hastalığı (AD) ve diğer nörodejeneratif bozukluklarda, oligodendroglial başarısızlık yaygın bir erken patolojik özelliktir, ancak özellikle beynin gri maddesinde hastalık gelişimine ve ilerlemesine nasıl katkıda bulunduğu büyük ölçüde bilinmemektedir. Oligodendrosit soy hücrelerinin işlev bozukluğu, miyelinasyondaki eksiklikler ve oligodendrosit öncü hücrelerinin (OPC'ler) bozulmuş kendini yenilemesi ile işaretlenmiştir. Bu iki kusur, en azından kısmen, patolojinin birikmesi boyunca nöron ve oligodendrositler arasındaki etkileşimlerin bozulmasından kaynaklanır. OPC'ler, CNS gelişimi sırasında miyelinizan oligodendrositlere yol açar. Olgun beyin korteksinde, OPC'ler başlıca proliferatif hücrelerdir (toplam beyin hücrelerinin ~% 5'ini oluşturur) ve yeni miyelin oluşumunu nöral aktiviteye bağımlı bir şekilde kontrol eder. Bu tür nöron-oligodendrosit iletişimi, özellikle AD gibi nörodejeneratif koşullar bağlamında, uygun araçların eksikliği nedeniyle önemli ölçüde az çalışılmaktadır. Son yıllarda, grubumuz ve diğerleri, insan pluripotent kök hücrelerinden bireysel olarak fonksiyonel nöronlar ve oligodendrositler üretmek için mevcut protokolleri geliştirmek için önemli ilerlemeler kaydetmiştir. Bu yazıda, nöron-oligodendrosit bağlantılarını modellemek için bir ko-kültür sisteminin kurulması da dahil olmak üzere optimize edilmiş prosedürlerimizi açıklamaktayız. Açıklayıcı sonuçlarımız, OPC'lerin / oligodendrositlerin beyin amiloidozuna ve sinaps bütünlüğüne beklenmedik bir katkısını önermektedir ve bu metodolojinin AD araştırması için faydasını vurgulamaktadır. Bu indirgemeci yaklaşım, spesifik hetero-hücresel etkileşimleri beynin içindeki doğal karmaşıklıktan ayırmak için güçlü bir araçtır. Burada tanımladığımız protokollerin, nörodejenerasyonun patogenezinde oligodendroglial defektler üzerine gelecekteki çalışmaları kolaylaştırması beklenmektedir.

İnsan pluripotent kök hücrelerinden insan kaynaklı nöronların doğrudan üretimi Yeni başlayan insan pluripotent kök hücrelerinin, başarılı bir şekilde iN veya iOPC / iOL üretimi için yüksek derecede pluripotens sergilemesi çok önemlidir. Bu nedenle, hücreler bu makalede açıklanan indüksiyon protokollerinden herhangi birine başlamadan önce Oct4 ve SOX2 gibi spesifik belirteçler için boyanmalıdır (Şekil 1A). İnsan H1 hücreleri, Zhang ve ark. ...

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

Nörodejenerasyonda nöron-glial etkileşimler yetersiz araç ve yöntemler nedeniyle iyi anlaşılamamıştır. Burada, insan pluripotent kök hücrelerinden indüklenmiş nöronlar, oligodendrosit öncü hücreleri ve oligodendrositler elde etmek için optimize edilmiş protokolleri açıklıyoruz ve Alzheimer hastalığında hücre tipine özgü katkıları anlamada bu yöntemlerin değerlerine örnekler sunuyoruz.

Nöron-Oligodendrosit Etkileşimlerini Modellemek için Pluripotent Kök Hücrelerden İnsan Nöronlarının ve Oligodendrositlerin Üretilmesi

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

Atılan İnsan Fetal Kortikal Doku Sinir Kök Hücre Üretimi

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

Küçük Molekül İndüksiyon Pluripotent İnsan Embriyonik Kök Hücreleri İnsan Nöronal atalarıdır ve Nöronlar Verimli türetilmesi

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

İnsan Pluripotent Kök Hücreler Telencephalic glutamaterjik Nöron Şartnamesi

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

İnsan Nöral Agregalarının bir cGMP uygulanabilir Açılım Yöntemi kök hücrelerin pluripotent kök hücreleri veya Fetal Beyin dokusundan türetilen

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

İnsan Pluripotent dan Sinir Crest Progenitör Hücreler besleyici ücretsiz Derivasyon Kök Hücre

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

İnsan Pluripotent Kök Hücreleri dopaminerjik Nöron Farklılaşma Yönetmen

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

İnsan yetişkin cilt fibroblastlar indüklenen Neurons yüksek verimli kültürünün basit üretimi

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

Neuritogenez ve Sinaps Matürasyonu yüksek içerikli Tarama için İnsan Pluripotent Kök Hücre Kaynaklı Nöronların Post-differentiation Rekaplamag

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

Insan kaynaklı pluripotent kök hücrelerinin (iPSCs), PiggyBac vektörler kullanarak fonksiyonel spinal ve kranial motor nöronlara dönüşümü

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

Oligodendrocytes ve Oligodendrocyte-Ko-Kültür Deneyleri için Şartlı Orta Üretimi

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