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Single-cell derived clones from human adipose stem cells present different immunomodulatory properties.

ABSTRACT: Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, 5 single cell clones were isolated (1.X and 3.X) from 2 volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1.10 and 1.22 expressed the lowest amounts, while clones 3.10 and 3.5 expressed more CD105 than the rest and clone 1.7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of IL-6 and low to moderate levels of IL-8. Furthermore, clone 3.X produced the highest amounts of pro-inflammatory cytokines such as IL-1β, while clones 1.10 and 1.22 highly expressed IL-4 and IL-5. These differences can be explained in part by the distinct DNA promoter methylation profile exhibited by the clones. The results of this work indicates that this stem cell population is heterogeneous in its secretion profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies. Total DNA isolated by standard procedures from different clones of human dipose mesenchymal stem cells (hMSC)

ORGANISM(S): Homo sapiens

SUBMITTER: Agustin Fernandez

PROVIDER: E-GEOD-44222 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Single-cell derived clones from human adipose stem cells present different immunomodulatory properties.

Project description:Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, 5 single cell clones were isolated (1.X and 3.X) from 2 volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1.10 and 1.22 expressed the lowest amounts, while clones 3.10 and 3.5 expressed more CD105 than the rest and clone 1.7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of IL-6 and low to moderate levels of IL-8. Furthermore, clone 3.X produced the highest amounts of pro-inflammatory cytokines such as IL-1β, while clones 1.10 and 1.22 highly expressed IL-4 and IL-5. These differences can be explained in part by the distinct DNA promoter methylation profile exhibited by the clones. The results of this work indicates that this stem cell population is heterogeneous in its secretion profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies.

2014-03-03 | GSE44222 | GEO

Cell of origin epigenetic priming determines susceptibility to Tet2 mutation

Project description:Hematopoietic stem cell (HSC) mutations can result in clonal hematopoiesis (CH) with heterogeneous clinical outcomes. Here, we investigated how the cell state preceding Tet2 mutation impacts the pre-malignant phenotype. Using an inducible system for clonal analysis of myeloid progenitors, we found that the epigenetic features of clones at similar differentiation status were highly heterogeneous and functionally responded differently to Tet2 mutation. Cell differentiation stage also influenced Tet2 mutation response indicating that the cell of origin's epigenome modulates clone-specific behaviors in CH. Molecular features associated with higher risk outcomes include Sox4 that sensitized cells to Tet2 inactivation, inducing dedifferentiation, altered metabolism and increasing the in vivo clonal output of mutant cells, as confirmed in primary GMP and HSC models. Our findings validate the hypothesis that epigenetic features can predispose specific clones for dominance, explaining why identical genetic mutations can result in different phenotypes.

1970-01-01 | MTBLS9914 | MetaboLights

LFQP of lysates from wild-type, RAB18-, TBC1D20-, RAB3GAP1- and RAB3GAP2-null RPE1 cells

Project description:These data are from two wild-type RPE1 clones (WT11, WT20), a RAB18-null clone, a TBC1D20-null clone, a RAB3GAP1-null clone and a RAB3GAP2-null clone, each analysed in triplicate.

2024-01-26 | PXD016326 | Pride

Daphnia magna CRISPR/Cas9 TRH gene knock downs transciptomic profilling

Project description:Transcriptomic profilling of 4 daphnia magna clones. One laboratory clone (Clone F +/+), one heterozygotic Clone (Clone 13 +/-) , two homozygotic Clones (Clone 16 and 17 -/-)

2018-07-31 | GSE101858 | GEO

Clonal Immortalized Human Glial Cell Lines Support Varying Levels of JC Virus Infection due to Differences in Cellular Gene Expression

Project description:JC virus (JCV) is a ubiquitous human polyomavirus that causes the demyelinating disease Progressive Multifocal Leukoencephalopathy (PML). JCV replicates in limited cell types in culture, predominantly in human glial cells. Thus, productive JCV infection is an indicator of the host cell transcription environment. Following introduction of a replication defective SV40 mutant that expressed large T protein into a heterogeneous culture of human fetal brain cells, multiple phenotypes became immortalized (SVG cells). A subset of SVG cells could support JCV replication. This mixed culture was called SVG cells. In the current study, clonal cell lines were selected from the original SVG cell culture. The SVG-5F4 clone showed low levels of viral growth. The SVG-10B1 clone was highly permissive for JCV DNA replication and gene expression. Microarray analysis revealed that viral infection did not significantly change gene expression in these cells. More resistant 5F4 cells expressed high levels of transcription factors known to inhibit JCV transcription. Interestingly, 5F4 cells highly expressed RNA of markers of Bergman or radial glia and 10B1 cells had high expression of markers of immature glial cells and activation of transcription regulators important for stem/progenitor cell self-renewal. These SVG-derived clonal cell lines provide a biologically relevant model to investigate cell type differences in JCV host range and pathogenesis, as well as neural development. 13 Human samples: 3 SVG 10B1 clones 14 days post mock-infection, 3 SVG 10B1 clones 14 days post JCV Mad-4 strain infection, 3 SVG 5F4 clones 14 days post mock-infection, 4 SVG 5F4 clones 14 days post JCV Mad-4 strain infection.

2013-10-17 | E-GEOD-45639 | biostudies-arrayexpress

Gene expression analysis of human fallopian tube stem cells and air-liquid interface differentiation

Project description:If the fallopian tube is the origin of serous cancer, one possible mechanism for the evolution of cancer is a dysregulation of indigenous stem cells. We therefore set out to clone the stem cells of the human fallopian tube using methods to clone columnar epithelial stem cells such as human intestinal stem cells. Using this method, we were able to generate clones of fallopian tube stem cells that contain many small, undifferentiated cells. These stem cell clones show strong and consistent staining with markers of fallopian tube epithelial cells (PAX8). We also established an air-liquid interface culture system to differentiate fallopian tube stem cell to both ciliated cells and non-ciliated cells.

2015-09-29 | GSE69453 | GEO

Epigenetic Memory Underlies Cell Autonomous Heterogeneous Behavior of Hematopoietic Stem Cells

Project description:Stem cells determine homeostasis and repair of many tissues and are increasingly recognized as functionally heterogeneous. To define the extent of and molecular basis for heterogeneity we overlaid functional, transcriptional and epigenetic attributes of hematopoietic stem cells (HSCs) at a clonal level using endogenous fluorescent tagging. Endogenous HSC had clone-specific functional attributes in vivo. The intra-clonal behaviors were highly stereotypic, conserved under the stress of transplantation, inflammation and genotoxic injury, and associated with distinctive transcriptional, DNA methylation and chromatin accessibility patterns. Further, HSC function corresponded to epigenetic configuration but not always to transcriptional state. Therefore, hematopoiesis under homeostatic and stress conditions represents the integrated action of highly heterogeneous clones of HSC with epigenetically scripted behaviors. This high degree of epigenetically driven cell autonomy among HSCs implies that refinement of the concepts of stem cell plasticity and of the stem cell niche are warranted.

2016-11-17 | GSE87527 | GEO

A class II-restricted chlamydia-specific CD8ɣ13 T cell clone protects the genital tract during Chlamydia muridarum infection

Project description:The T cell response to Chlamydia genital tract infections in humans and mice is unusual in that the majority of antigen-specific CD8 T cells are not restricted by HLA/MHC class I and therefore have been referred to as “unrestricted” or “atypical”. We previously reported that a subset of unrestricted murine Chlamydia-specific CD8 T cells had an unusual cytokine polarization pattern that included IFN-ɣ and IL-13. For this report, we investigated the transcriptome of Chlamydia-specific CD8ɣ13 T cells, comparing them to Chlamydia-specific multifunctional Tc1 clones using gene expression micro array analysis. The molecular study revealed that CD8ɣ13 polarization included IL-5 in addition to IFN-γ and IL-13. Adoptive transfer studies were performed with Tc1 clone and CD8ɣ13 T cell clones to determine whether either influenced bacterial clearance or immunopathology during Chlamydia muridarum (Cm) genital tract infections. To our surprise, an adoptively transferred CD8ɣ13 T cell clone was remarkably proficient at preventing chlamydia immunopathology while the multifunctional Tc1 clone did not enhance clearance or significantly protect from immunopathology. Mapping studies with MHC class I- and class II-deficient splenocytes showed our previously published Chlamydia-specific CD8 T cell clones are MHC class II-restricted. MHC class II-restricted CD8 T cells may play important roles in protection from intracellular pathogens that limit class I antigen presentation or deplete the CD4 T cell compartment.

2020-05-21 | GSE140213 | GEO

Comparative membrane protein composition of Trichomonas gallinae clones with different virulence

Project description:This project compares the composition of proteins present in the cellular membrane and organelles membranes of two clones of Trichomonas gallinae that differed in virulence. The virulence was first tested in cell cultures. Two out of seven clones were selected according to their difference in virulence: the clone with the highest initial score of cytopatic effect (CPE) and the clone with the lowest initial score of CPE. The experiment was done using a label free approach.

2019-10-10 | PXD013115 | Pride

Gene expression profiling of Nup98HoxA9 expressing ES cells

Project description:To know whether the Nup98-HoxA9 affects global gene expression, we performed DNA microarray analysis using the following four clones: two independent Nup98-HoxA9 ES clones (clone#1 and clone#9), parental ES cells, and HoxA9-Ct ES clone.

2016-01-04 | GSE67967 | GEO

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