Gene expression profiles of human iPSC-derived microglia in monoculture and in co-culture with isogenic iPSC-derived neural cells
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE243370 | GEO | 2024/08/15
REPOSITORIES: GEO
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Project description:Gene expression profiles of human iPSC-derived microglia in monoculture and in co-culture with isogenic iPSC-derived neural cells
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Project description:Motor neuron diseases such as amyotrophic lateral sclerosis are primarily characterized by motor neuron degeneration with additional involvement of non-neuronal cells, in particular, microglia. In previous work, we have established protocols for the differentiation of iPSC-derived spinal motor neurons and microglia. Here, we combine both cell lineages and establish a novel co-culture of iPSC-derived spinal motor neurons and microglia, which is compatible with motor neuron identity and function. Co-cultured microglia express key identity markers and transcriptomically resemble primary human microglia, have highly dynamic ramifications, are phagocytically competent, release relevant cytokines and respond to stimulation. Further, they express key amyotrophic lateral sclerosis-associated genes and release disease-relevant biomarkers. This novel and authentic human model system facilitates the study of physiological motor neuron-microglia crosstalk and will allow the investigation of non-cell-autonomous phenotypes in motor neuron diseases such as amyotrophic lateral sclerosis.
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Project description:There is increasing genetic evidence for the role of microglia in neurodegenerative diseases, including Alzheimer's, Parkinson's, and motor neuron disease. Therefore, there is a need to generate authentic in vitro models to study human microglial physiology. Various methods have been developed using human induced Pluripotent Stem Cells (iPSC) to generate microglia, however, systematic approaches to identify which media components are actually essential for functional microglia are mostly lacking. Here, we systematically assess medium components, coatings, and growth factors required for iPSC differentiation to microglia. Using single-cell RNA sequencing, qPCR, and functional assays, with validation across two labs, we have identified several medium components from previous protocols that are redundant and do not contribute to microglial identity. We provide an optimised, defined medium which produces both transcriptionally and functionally relevant microglia for modelling microglial physiology in neuroinflammation and for drug discovery.
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Project description:To understand the behaviour in terms of special genome components that are expressed by L.monocytogenes in the presence of another bacterium as may be the condition in its natural environments was our objective, for which we have used microarray gene expression at different time intervals of growth from 4hrs to 24 hrs. Expression of L.monocytogenes as co-cultures, both in broth culture state and biofilm state were differentiated to that of 24hrs pure broth culture. Also genes regulated for and during biofilm formation as pure cultures were identified with comparision to 24hrs pure broth culture. Distinguishing features with notable variation in all of the three sample sets as L.monocytogenes in pure culture biofilm, co-culture broth and co-culture biofilm were observable. Genes that are specifically up-regulated at each of growth condition and time interval were identified, genes regulated with ascending and descending patterns in time were also noticable. These variation in the gene expression gives an insight into the alterations in the biosynthetic and metabolic pathways under different states of growth
2012-01-15 | GSE27936 | GEO
Project description:In this study, we constructed a tissue-engineered DRG-CSC (corneal stromal cells) co-cultured model (DS model), a CSC monoculture model (S model), and a DRG monoculture model (D model). Chicken DRGs were extracted from E7-E10 chicken embryo. Human CSCs were extracted in stromal lenticules from small incision lenticule extraction (SMILE). Transcriptional profiling of DRG in the DS model (DS-D), DRG in the D model(D-D), CSCs in the DS model (DS-S), CSCs in the S model (S-S) was analyzed. After 7 days of culture, RNA of each group was extracted.
2021-06-29 | GSE179020 | GEO
Project description:Expression data comparing the effect of I1061T NPC1 mutation and anti-CD22 treatment in iPSC-derived microglia like cells treated with NPC patient CSF.
2021-09-18 | GSE184243 | GEO
Project description:Human induced pluripotent stem cells (iPSCs), together with 21st century cell culture methods, have the potential to better model human physiology with applications in toxicology, disease modeling, and the study of host-pathogen interactions. Several models of the human brain have been developed recently, demonstrating cell-cell interactions of multiple cell types with physiologically relevant 3D structures. Most current models, however, lack the ability to represent the inflammatory response in the brain because they do not include a microglial cell population. Microglia, the resident immunocompetent phagocytes in the central nervous system (CNS), are not only important in the inflammatory response and pathogenesis; they also function in normal brain development, strengthen neuronal connections through synaptic pruning, and are involved in oligodendrocyte and neuronal survival. Here, we have successfully introduced a population of human microglia into 3D human iPSC-derived brain spheres (BrainSpheres, BS) through co-culturing cells of the Immortalized Human Microglia - SV40 cell line with the BS model (?BS). We detected an inflammatory response to lipopolysaccharides (LPS) and flavivirus infection, which was only elicited in the model when microglial cells were present. A concentration of 20 ng/mL of LPS increased gene expression of the inflammatory cytokines interleukin-6 (IL-6), IL-10, and IL-1?, with maximum expression at 6-12 h post-exposure. Increased expression of the IL-6, IL-1?, tumor necrosis factor alpha (TNF-?), and chemokine (C-C motif) ligand 2 (CCL2) genes was observed in ?BS following infection with Zika and Dengue Virus, suggesting a stronger inflammatory response in the model when microglia were present than when only astrocyte, oligodendrocyte, and neuronal populations were represented. Microglia innately develop within cerebral organoids (Nature communications), our findings suggest that the ?BS model is more physiologically relevant and has potential applications in infectious disease and host-pathogen interactions research.
| S-EPMC6296317 | biostudies-literature
Project description:BackgroundWe previously reported that the pluripotent stem cells can differentiate into cardiomyocytes (CMs) by co-culture with neonatal CMs (NCMs) in vitro. However, the involving mechanism is not clear.MethodsMouse induced pluripotent stem cells (iPSCs) were cultured in hanging drops to form embryoid bodies (EBs) and to induce myocardial differentiation. Co-culture of EBs and NCMs was established in a transwell insert system, while EBs grown alone in the wells were used as controls.ResultsCo-culture with NCMs markedly increased the generation of functional CMs from iPSCs. The focal adhesion kinase (FAK) phosphorylation, and c-Jun N-terminal kinase (JNK) phosphorylation in co-culture were higher than that in EBs grown alone. Treating FAK small interfering RNA (FAK siRNA) or specific inhibitor for JNK (SP600125) to iPSCs significantly reduced the phosphorylation of JNK and the expressions of Mef2c and Bcl-2. The expressions of cTnT and MLC-2V were also decreased. Our results revealed that co-culture with NCMs significantly enhance the differentiation ability of iPSCs by increasing Mef2c and Bcl-2 expressions concomitantly with a marked augment on cell proliferation through JNK signaling pathways.ConclusionsThese findings indicated that co-culture of EBs with NCMs induces genes expressed in a mature pattern and stimulates the proliferation of iPSC-derived CMs (iPS-CMs) by activating FAK/JNK signaling.
| S-EPMC4855360 | biostudies-literature
Project description:To understand the behaviour in terms of special genome components that are expressed by L.monocytogenes in the presence of another bacterium as may be the condition in its natural environments was our objective, for which we have used microarray gene expression at different time intervals of growth from 4hrs to 24 hrs. Expression of L.monocytogenes as co-cultures, both in broth culture state and biofilm state were differentiated to that of 24hrs pure broth culture. Also genes regulated for and during biofilm formation as pure cultures were identified with comparision to 24hrs pure broth culture. Distinguishing features with notable variation in all of the three sample sets as L.monocytogenes in pure culture biofilm, co-culture broth and co-culture biofilm were observable. Genes that are specifically up-regulated at each of growth condition and time interval were identified, genes regulated with ascending and descending patterns in time were also noticable. These variation in the gene expression gives an insight into the alterations in the biosynthetic and metabolic pathways under different states of growth Agilent one-color experiment,Organism: Listeria monocytogenes , Genotypic Technology Pvt. Ltd. designed Custom Microarray Listeria monocytogenes 8x15k (Agilent-30831) , Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)
2012-01-15 | E-GEOD-27936 | biostudies-arrayexpress
Project description:Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease in which upper and lower motor neuron loss is the primary phenotype, leading to muscle weakness and wasting, respiratory failure, and death. Although a portion of ALS cases are linked to one of over 50 unique genes, the vast majority of cases are sporadic in nature. However, the mechanisms underlying the motor neuron loss in either familial or sporadic ALS are not entirely clear. Here, we used induced pluripotent stem cells derived from a set of identical twin brothers discordant for ALS to assess the role of astrocytes and microglia on the expression and accumulation of neurofilament proteins in motor neurons. We found that motor neurons derived from the affected twin which exhibited increased transcript levels of all three neurofilament isoforms and increased expression of phosphorylated neurofilament puncta. We further found that treatment of the motor neurons with astrocyte-conditioned medium and microglial-conditioned medium significantly impacted neurofilament deposition. Together, these data suggest that glial-secreted factors can alter neurofilament pathology in ALS iPSC-derived motor neurons.
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