<i>In vitro</i> myelin formation using embryonic stem cells

Kerman, Bilal E.; Kim, Hyung Joon; Padmanabhan, Krishnan; Mei, Arianna; Georges, Shereen; Joens, Matthew S.; Fitzpatrick, James A.J.; Jappelli, Roberto; Chandross, Karen J.; August, Paul R.; Gage, Fred H.

doi:10.1242/dev.116517

Cited by 83 publications

(92 citation statements)

References 49 publications

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“…The protocol to obtain a myelinating stem cell-derived co-culture has recently been described [25]. In this study, the functionality of stem cell-derived oligodendrocytes in terms of myelination has been confirmed by demonstrating the formation of physiological myelin on an ultrastructural level.…”

Section: Methodsmentioning

confidence: 64%

“…Briefly, E14Tg2a mouse embryonic stem cells were differentiated into cortical neurons [17]. To obtain mature oligodendrocytes, neural progenitor cells were derived from mouse D3 embryonic stem cells as described elsewhere [34] and underwent a two-step differentiation/maturation protocol [25]. OPCs were differentiated from neural progenitor cells using differentiation medium (DMEM/F12-GlutaMax, N2, B27, 10ng/ml insulin-like growth factor-1, 10 ng/ml PDGF-aa) for 7 days before switching to maturation medium for another 7 days (DMEM/F12-GlutaMax, N2, B27, 10 ng/ml ciliary neurotrophic factor, 5 ng/ml neurotrophin-3, 40 ng/ml triiodothyronine (T3)).…”

Section: Methodsmentioning

confidence: 99%

“…Myelinating co-cultures and D3 embryonic stem cell-derived oligodendrocytes were stained as recently described [25]. The following primary antibodies were used in combination for overnight incubation: chicken anti Gfp (Aves Labs, GFP-1020; 1:250), rat anti Mbp (AbD Serotec; MCA409S; 1:50), mouse anti beta-III-tubulin (Tuj1, Covance, MMS-435P; 1:1000), rat anti human α-synuclein (15G7, ENZO; ALX-804-258; 1:200), rabbit anti Pdgfrα (Santa Cruz; sc-338; 1:400), rabbit anti cleaved Caspase 3 (Asp175; Cell Signaling; #9661; 1:500).…”

Section: Methodsmentioning

confidence: 99%

“…Myelination in the oligodendrocyte-neuron co-culture was semi-automatically quantified using the Computer-assisted Evaluation of Myelin formation (CEM) tool [25]. Myelination was assessed by analyzing individual oligodendrocytes.…”

Section: Methodsmentioning

confidence: 99%

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α-Synuclein-induced myelination deficit defines a novel interventional target for multiple system atrophy

et al. 2016

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Multiple system atrophy (MSA) is a rare atypical parkinsonian disorder characterized by a rapidly progressing clinical course and at present without any efficient therapy. Neuropathologically, myelin loss and neurodegeneration are associated with α-synuclein accumulation in oligodendrocytes, but underlying pathomechanisms are poorly understood. Here, we analyzed the impact of oligodendrocytic α-synuclein on the formation of myelin sheaths in order to define a potential interventional target for MSA. Post-mortem analyses of MSA patients and controls were performed to quantify myelin and oligodendrocyte numbers. As pre-clinical models, we used transgenic MSA mice, a myelinating stem cell-derived oligodendrocyte-neuron co-culture, and primary oligodendrocytes to determine functional consequences of oligodendrocytic α-synuclein overexpression on myelination. We detected myelin loss accompanied by preserved or even increased numbers of oligodendrocytes in post-mortem MSA brains or transgenic mouse forebrains, respectively, indicating an oligodendrocytic dysfunction in myelin formation. Corroborating this observation, overexpression of α-synuclein in primary and stem cell-derived oligodendrocytes severely impaired myelin formation, defining a novel α-synuclein-linked pathomechanism in MSA. We used the pro-myelinating activity of the muscarinic acetylcholine receptor antagonist benztropine to analyze the reversibility of the myelination deficit. Transcriptome profiling of primary pre-myelinating oligodendrocytes demonstrated that benztropine readjusts myelination-related processes such as cholesterol and membrane biogenesis, being compromised by oligodendrocytic α-synuclein. Additionally, benztropine restored the α-synuclein-induced myelination deficit of stem cell-derived oligodendrocytes. Strikingly, benztropine also ameliorated the myelin deficit in transgenic MSA mice, resulting in a prevention of neuronal cell loss. In conclusion, this study defines the α-synuclein-induced myelination deficit as a novel and crucial pathomechanism in MSA. Importantly, the reversible nature of this oligodendrocytic dysfunction opens a novel avenue for an intervention in MSA.

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Section: Methodsmentioning

confidence: 64%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

α-Synuclein-induced myelination deficit defines a novel interventional target for multiple system atrophy

et al. 2016

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“…Existing protocols for generating myelinating oligodendrocytes in vitro from hPSCs are extremely inefficient. Current protocols use a combination of factors to drive initial neuralization, followed by the addition of oligoglia-inducing factors (such as, PDGFα, neurotrophin 3 (NT3) and insulin-like growth factors (IGFs)) to generate oligodendrocyte progenitor cells (OPCs) 39,75,76 . Sufficient myelination is typically only achieved following in vivo transplantation 77 .…”

Section: Directed Hpsc Differentiationmentioning

confidence: 99%

Evaluating cell reprogramming, differentiation and conversion technologies in neuroscience

et al. 2016

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The scarcity of live human brain cells for experimental access has for a long time limited our ability to study complex human neurological disorders and elucidate basic neuroscientific mechanisms. A decade ago, the development of methods to reprogramme somatic human cells into induced pluripotent stem cells enabled the in vitro generation of a wide range of neural cells from virtually any human individual. The growth of methods to generate more robust and defined neural cell types through reprogramming and direct conversion into induced neurons has led to the establishment of various human reprogramming-based neural disease models.

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Controlling Differentiation of Stem Cells for Developing Personalized Organ‐on‐Chip Platforms

Geraili

Jafari

Hassani

et al. 2017

Adv Healthcare Materials

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Organ‐on‐chip (OOC) platforms have attracted attentions of pharmaceutical companies as powerful tools for screening of existing drugs and development of new drug candidates. OOCs have primarily used human cell lines or primary cells to develop biomimetic tissue models. However, the ability of human stem cells in unlimited self‐renewal and differentiation into multiple lineages has made them attractive for OOCs. The microfluidic technology has enabled precise control of stem cell differentiation using soluble factors, biophysical cues, and electromagnetic signals. This study discusses different tissue‐ and organ‐on‐chip platforms (i.e., skin, brain, blood–brain barrier, bone marrow, heart, liver, lung, tumor, and vascular), with an emphasis on the critical role of stem cells in the synthesis of complex tissues. This study further recaps the design, fabrication, high‐throughput performance, and improved functionality of stem‐cell‐based OOCs, technical challenges, obstacles against implementing their potential applications, and future perspectives related to different experimental platforms.

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In vitro myelin formation using embryonic stem cells

Cited by 83 publications

References 49 publications

α-Synuclein-induced myelination deficit defines a novel interventional target for multiple system atrophy

α-Synuclein-induced myelination deficit defines a novel interventional target for multiple system atrophy

Evaluating cell reprogramming, differentiation and conversion technologies in neuroscience

Controlling Differentiation of Stem Cells for Developing Personalized Organ‐on‐Chip Platforms

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