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Rapid and efficient generation of myelinating oligodendrocytes from human induced pluripotent stem cells using a combination of three transcription factors [hiPSC]

ABSTRACT: We demonstrate that the induction of three transcription factors (SOX10, OLIG2, NKX6.2) in hiPSC-derived neural progenitor cells (hiPSC-NPC) is sufficient to rapidly generate O4+ oligodendrocytes with an efficiency of 60 to 70% within 28 days.

ORGANISM(S): Homo sapiens

PROVIDER: GSE79912 | GEO | 2017/03/06

SECONDARY ACCESSION(S): PRJNA317361

REPOSITORIES: GEO

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Rapid and efficient generation of myelinating oligodendrocytes from human induced pluripotent stem cells using a combination of three transcription factors [brain]

Project description:We demonstrate that the induction of three transcription factors (SOX10, OLIG2, NKX6.2) in hiPSC-derived neural progenitor cells (hiPSC-NPC) is sufficient to rapidly generate O4+ oligodendrocytes with an efficiency of 60 to 70% within 28 days.

2017-03-06 | GSE79913 | GEO

RNA-seq of in vitro reprogrammed mouse pericytes to oligodendrocytes

Project description:Direct conversion of pericytes (PCs) or mouse embryonic fibroblasts (MEFs) into induced oligodendrocytes (iOPCs) by ectopic expression of Olig2, Sox10 and Nkx6.2 was assessed by transcriptome profiling (RNA-seq). Samples were collected before and after direct conversion (for PCs at 3 different time points/passages - p5, p15 and p25)

2021-03-07 | E-MTAB-8453 | biostudies-arrayexpress

Rapid differentiation of hiPSCs into functional oligodendrocytes using an OLIG2 synthetic modified messenger RNARapid differentiation of hiPSCs into functional oligodendrocytes using an OLIG2 synthetic modified messenger RNA

Project description:We show that a synthetic modified messenger RNA (smRNA)-based reprogramming method that leads to the generation of transgene-free OLs has been developed. An smRNA encoding a modified form of OLIG2, a key TF in OL development, in which the serine 147 phosphorylation site is replaced with alanine, OLIG2S147A, is designed to reprogram hiPSCs into OLs. We demonstrate that repeated administration of the smRNA encoding OLIG2 S147A lead to higher and more stable protein expression. Using the single-mutant OLIG2 smRNA morphogen, we establish a 6-day smRNA transfection protocol, and glial induction lead to rapid NG2+ OL progenitor cell (OPC) generation (> 70% purity) from hiPSC-derived neural progenitor cells (NPCs). The smRNA-induced NG2+ OPCs can mature into functional OLs in vitro and promote remyelination in vivo. Proteomic analysis of OLIG2-binding proteins indicates that OLIG2 is bound by the heat shock protein 70 (HSP70) complex. The HSP70 complex is bound more strongly to OLIG2 with the modified phosphorylation site than to wild-type OLIG2.

2023-03-10 | PXD036975 | Pride

Rapid and Efficient Generation of Myelinating Human Oligodendrocytes in Organoids.

Project description:Human stem cell derived brain organoids are increasingly gaining attention as an ideal model system for investigating neurological diseases, particularly those that involve myelination defects. However, current protocols for generating brain organoids with sufficiently mature oligodendrocytes that deposit myelin on endogenously produced neurons are lengthy and complicated. Taking advantage of a human pluripotent stem cell line that reports on SOX10 expression, we developed a protocol that involves a 42 day exposure of neuroectoderm-derived organoids to a cocktail of growth factors and small molecules that collectively foster oligodendrocyte specification and survival. Importantly, the resulting day 42 brain organoids contain both myelinating oligodendrocytes, cortical neuronal cells and astrocytes. These oligodendrocyte brain organoids therefore constitute a valuable and tractable platform for functional neurogenomics and drug screening for white matter diseases.

| S-EPMC8010307 | biostudies-literature

RNA-seq, ATAC-seq and DNA-methylation array data of human oligodendrocyte-like cells derived from transdifferentiated fibroblasts

Project description:One-step reprogramming of human fibroblasts into oligodendrocyte-like cells. We characterized the expression, chromatin accessibility and methylation profile of human oligodendrocyte-like cells derived using a new protocol for the direct transdifferentiation of human fibroblasts into oligodendrocyte-like cells. Direct conversion of fibroblasts was induced by ectopic overexpression of SOX10, OLIG2 and NKX6.2. The aim of the experiments was to characterise the transcriptional response and associated chromatin changes following transdifferentiation. In addition, profiles of primary human oligodendrocytes were produced.

2021-03-22 | E-MTAB-7838 | biostudies-arrayexpress

ATAC-seq of in vitro reprogrammed mouse pericytes to oligodendrocytes

Project description:Direct conversion of pericytes (PCs) into induced oligodendrocytes (iOPCs) by ectopic expression of Olig2 and Sox10 (2F) was assessed by chromatin accessibility profiling (by ATAC-seq). Samples were collected before and after direct conversion (at 3 different time points/passages - p5, p15 and p25).

2021-03-07 | E-MTAB-8452 | biostudies-arrayexpress

Generation of oligodendroglial cells by direct lineage conversion

Project description:We report the generation of induced oligodendrocyte precursor cells (iOPCs) by direct lineage conversion. Forced expression of the three transcription factors Sox10, Olig2 and Zfp536 was sufficient to convert mouse and rat fibroblasts into iOPCs with morphologies and gene expression signatures that resemble OPCs. We compared the global gene expression pattern of iOPCs, fibroblasts, primary OPCs from the neonatal rat brain, and their differentiated progeny. We purified iOPCs by O4 immunopanning three weeks after infection and extracted total RNA. Acutely isolated rat cortical OPCs were either used directly for RNA extraction or expanded in mitogen-containing media for 24h before switching into differentiation medium, lacking PDGF/NT-3 and containing T3. Cells were harvested for microarray analysis 3 and 6 days after induction of differentiation.

2013-04-07 | E-GEOD-40421 | biostudies-arrayexpress

Remarkable Stability of Myelinating Oligodendrocytes in Mice.

Project description:New myelin-forming oligodendrocytes (OLs) are generated in the mouse central nervous system during adulthood. These adult-born OLs might augment the existing population, contributing to neural plasticity, or else replace OLs that die in use (turnover). To distinguish between these alternatives, we induced genetic labeling of mature myelinating OLs in young adult mice and tracked their subsequent survival. OL survival rates were region dependent, being higher in corpus callosum (∼90% survival over 20 months) and motor cortex (∼70% survival) than in corticospinal tract or optic nerve (50%-60% survival). Survival rates over the first 8 months were 90%-100% in all regions except the optic nerve. In the corpus callosum, new OLs accumulate during young adulthood and are therefore likely to participate in adaptive myelination. We also found that the number of myelin internodes maintained by individual cortical OLs is stable for at least 8 months but declines ∼12% in the following year.

| S-EPMC5643547 | biostudies-literature

Differentiation and maturation of oligodendrocytes in human three-dimensional neural cultures

Project description:We report a method for deriving oligodendrocyte lineage cells from human pluripotent stem cells (hPSCs) in three-dimensional (3D) culture called human oligodendrocyte spheroids (hOLS). To characterize oligodendrocyte-lineage cells in hOLS, we isolated O4+ cells by immunopanning and performed deep single cell RNA sequencing. We sequenced 295 cells and compared their profiles to unsorted cells isolated from primary human fetal cortex, primary human adult cortex, and hCS. Clustering of all cells using the t-distributed stochastic neighbor embedding (tSNE) approach revealed a distinct populations of SOX10+ oligodendrocytes, within which the O4+ cells derived from hOLS clustered most closely to oligodendrocyte progenitor cells (OPCs) and mature oligodendrocytes from the primary human adult cortical tissue. Additionally, subpopulations of OPCs, newly formed oligodendrocytes, and myelinating oligodendrocytes derived were observed in the hOLS-derived cluster. To further assess the state of oligodendrocyte-lineage cells in hOLS, we performed a Monocle analysis which revealed a spectrum of oligodendrocyte-lineage stages in hOLS ranging from dividing cells that closely resembled primary OPCs to mature cells that closely resembled primary oligodendrocytes.

2018-12-31 | GSE115011 | GEO

Phenotypic Correction of Oligodendrocytes by Concurrent Direct Reprogramming and Gene Replacement of Fibroblasts

Project description:Transcription factor-mediated direct reprogramming technologies that can convert one differentiated somatic cell to another, without an intermediate pluripotent stem cell stage, provide a new mechanism to access somatic cell types for disease modeling and possible therapeutic application. Previous efforts have focused on wild-type cells harboring transgenic reporters to signal successful reprogramming events. Outstanding questions remain of whether cells from disease backgrounds without transgenic reporters are capable of direct reprogramming and whether they phenocopy disease pathology. Here we demonstrate that fibroblasts from the shiverer mouse model of myelin disease can be directly reprogrammed to induced oligodendrocyte progenitor cells (iOPCs) by the expression of Sox10, Olig2, and Nkx6.2 without the aid of transgenic selection reporters. Shiverer mice harbor a homozygous ~20-kilobase deletion in the Myelin Basic Protein (Mbp) gene and this substantial deletion leads to abnormal oligodendrocytes resulting in hypomyelination and decreased lifespan. Shiverer iOPCs efficiently differentiated to induced oligodendrocytes (iOLs) which recapitulated the known disease phenotype of impaired axonal ensheathment and myelination capacity. Combining direct reprogramming and genetic supplementation of a minigene carrying regulatory elements and an open reading frame of MBP, restored MBP expression in differentiated iOLs and their ability to track along and ensheath microfibers in an in vitro assay of myelination capacity. Collectively these results show that iOPCs provide a rapid and accessible system to accurately model myelin disease pathology and provide promise for future therapeutic advancement for genetic myelin disorders.

2020-05-02 | GSE98491 | GEO

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