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ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Homo sapiens

PROVIDER: GSE219026 | GEO | 2023/03/01

REPOSITORIES: GEO

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Effect of the Enrichment in c-Kit Stem Cell Potential of Foetal Human Amniotic Fluid Cells: Characterization from Single Cell Analysis to the Secretome Content.

Project description:Human amniotic fluid cells (hAFSCs) are a fascinating foetal cell-type that have important stem cell characteristics; however, they are a heterogeneous population that ranges from totally differentiated or progenitor cells to highly multipotent stem cells. There is no single approach to isolating the stem cell component, but the selection of a subpopulation of hAFSCs expressing c-Kit is widely employed, while a deep characterization of the two populations is still lacking. Here we performed single-cell and bulk RNAseq analysis to compare the gene expression profiles of adherent amniotic fluid cells and their subpopulation c-Kit+. Information deriving from this high throughput technology on the transcriptome was then confirmed for specific targets with protein expression experiments and functional analysis. In particular, transcriptome profiling identified changes in cellular distribution among the different clusters that correlated with significant differential expression in pathways related to stemness, proliferation, and cell cycle checkpoints. These differences were validated by RT-PCR, immunofluorescence, WB, and cell cycle assays. Interestingly, the two populations produced secretomes with different immune-modulating and pro-regenerative potentials. Indeed, the presence of TGFβ, HGF, IDO was higher in EVs deriving from c-Kit+ cells, unlike IL-6. These results suggest the existence of deep intra-population differences that can influence the stemness profile of hAFSCs. This study represents a proof-of-concept of the importance of selecting c-Kit positive fractions with higher potential in regenerative medicine applications.

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ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT [scRNA-Seq]

Project description:Human amniotic fluid cells (hAFSCs) are a fascinating foetal cell-type that have important stem cell characteristics, since described as broadly multipotent immature cells with high self-renewal and no tumorigenic properties. However, amniotic fluid contains a heterogeneous population of amniocytes, from totally differentiated or progenitor cells to highly multipotent stem cells. There is no single approach to isolating the stem cell component, but the selection of a subpopulation of hAFSCs expressing c-Kit (or CD117) is widely employed, while a deep characterization of the two populations is still lacking. Here we performed single-cell and bulk RNAseq analysis to compare the gene expression profiles of adherent amniotic fluid cells and their subpopulation c-Kit+. Information deriving from this high throughput technology on the transcriptome was then confirmed for specific targets with protein expression experiments and functional analysis.

2023-03-01 | GSE219023 | GEO

ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT [Bulk RNA-Seq]

2023-03-01 | GSE219024 | GEO

ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT

Project description:ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT

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ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT [scRNA-Seq]

Project description:ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT [scRNA-Seq]

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ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT [Bulk RNA-Seq]

Project description:ENRICHMENT IN C-KIT IMPROVES STEM CELL POTENTIAL OF FOETAL HUMAN AMNIOTIC FLUID CELLS: CHARACTERIZATION FROM SINGLE CELL ANALYSIS TO THE SECRETOME EFFECT [Bulk RNA-Seq]

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The human amniotic fluid stem cell secretome effectively counteracts doxorubicin-induced cardiotoxicity.

Project description:The anthracycline doxorubicin (Dox) is widely used in oncology, but it may cause a cardiomyopathy with bleak prognosis that cannot be effectively prevented. The secretome of human amniotic fluid-derived stem cells (hAFS) has previously been demonstrated to significantly reduce ischemic cardiac damage. Here it is shown that, following hypoxic preconditioning, hAFS conditioned medium (hAFS-CM) antagonizes senescence and apoptosis of cardiomyocytes and cardiac progenitor cells, two major features of Dox cardiotoxicity. Mechanistic studies with mouse neonatal ventricular cardiomyocytes (mNVCM) reveal that hAFS-CM inhibition of Dox-elicited senescence and apoptosis is associated with decreased DNA damage, nuclear translocation of NF-kB, and upregulation of the NF-kB controlled genes, Il6 and Cxcl1, promoting mNVCM survival. Furthermore, hAFS-CM induces expression of the efflux transporter, Abcb1b, and Dox extrusion from mNVCM. The PI3K/Akt signaling cascade, upstream of NF-kB, is potently activated by hAFS-CM and pre-treatment with a PI3K inhibitor abrogates NF-kB accumulation into the nucleus, modulation of Il6, Cxcl1 and Abcb1b, and prevention of Dox-initiated senescence and apoptosis in response to hAFS-CM. These results support the concept that hAFS are a valuable source of cardioprotective factors and lay the foundations for the development of a stem cell-based paracrine treatment of chemotherapy-related cardiotoxicity.

| S-EPMC4956770 | biostudies-literature

The human amniotic fluid stem cell secretome counteracts doxorubicin-induced cardiotoxicity

Project description:The anthracycline, doxorubicin (Dox), is widely used in oncology, but it may it may cause a cardiomyopathy which has dismal prognosis and cannot be effectively prevented. The secretome of multipotent human amniotic fluid-derived stem cells (hAFS) has previously been demonstrated to reduce ischemic cardiac damage. Here, it is shown that the hAFS conditioned medium (hAFS-CM) antagonizes senescence and apoptosis of cardiomyocytes and cardiac progenitor cells, two major features of Dox cardiotoxicity. Mechanistic studies with primary mouse neonatal cardiomyocytes reveal that hAFS-CM inhibition of Dox-elicited senescence and apoptosis is paralleled by decreased DNA damage and is associated with nuclear translocation of NF-kB and upregulation of a set of genes controlled by NF-kB, namely Il6 and Cxcl1, which promote cardiomyocyte survival, and Cyp1b1 and Abcb1, which encode for proteins involved in Dox metabolism and efflux, respectively. The PI3K/Akt signaling cascade, upstream of NF-kB, is potently activated by the hAFS-CM and pre-treatment with a PI3K inhibitor abrogates NF-kB accumulation into the nucleus, modulation of its target genes, and prevention of Dox-initiated senescence and apoptosis in response to the hAFS-CM. This work may lay the ground for the development of a stem cell-based paracrine therapy of chemotherapy-related cardiotoxicity.

2016-10-30 | GSE74513 | GEO

Hypoxia-induced amniotic fluid stem cell secretome augments cardiomyocyte proliferation and enhances cardioprotective effects under hypoxic-ischemic conditions.

Project description:Secretome derived from human amniotic fluid stem cells (AFSC-S) is rich in soluble bioactive factors (SBF) and offers untapped therapeutic potential for regenerative medicine while avoiding putative cell-related complications. Characterization and optimal generation of AFSC-S remains challenging. We hypothesized that modulation of oxygen conditions during AFSC-S generation enriches SBF and confers enhanced regenerative and cardioprotective effects on cardiovascular cells. We collected secretome at 6-hourly intervals up to 30 h following incubation of AFSC in normoxic (21%O2, nAFSC-S) and hypoxic (1%O2, hAFSC-S) conditions. Proliferation of human adult cardiomyocytes (hCM) and umbilical cord endothelial cells (HUVEC) incubated with nAFSC-S or hAFSC-S were examined following culture in normoxia or hypoxia. Lower AFSC counts and richer protein content in AFSC-S were observed in hypoxia. Characterization of AFSC-S by multiplex immunoassay showed higher concentrations of pro-angiogenic and anti-inflammatory SBF. hCM demonstrated highest proliferation with 30h-hAFSC-S in hypoxic culture. The cardioprotective potential of concentrated 30h-hAFSC-S treatment was demonstrated in a myocardial ischemia-reperfusion injury mouse model by infarct size and cell apoptosis reduction and cell proliferation increase when compared to saline treatment controls. Thus, we project that hypoxic-generated AFSC-S, with higher pro-angiogenic and anti-inflammatory SBF, can be harnessed and refined for tailored regenerative applications in ischemic cardiovascular disease.

| S-EPMC7794288 | biostudies-literature

Supporting data on in vitro cardioprotective and proliferative paracrine effects by the human amniotic fluid stem cell secretome.

Project description:The data and information presented here refer to the research article entitled: "Reactivating endogenous mechanisms of cardiac regeneration via paracrine boosting with the human amniotic fluid stem cell secretome" (Balbi et al., 2019, Apr 04). This dataset illustrates the in vitro paracrine effect exerted by the human amniotic fluid stem cell secretome on rodent neonatal cardiomyocytes, human endothelial progenitors and different subsets of cardiac progenitor cells. Cytokine/chemokine profiling of the human amniotic fluid stem cell secretome is provided as well. This data can provide useful insights in regenerative medicine as demonstrating the in vitro cardioprotective and proliferative secretory paracrine potential of human fetal stem cells.

| S-EPMC6700494 | biostudies-literature

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