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Cultured human amniotic fluid-derived mesenchymal stromal cells [miRXplore data]

ABSTRACT: Human amniotic fluid-derived mesenchymal stromal cells (hAMSC) have become one of the main cell populations used in regenerative medicine and for the study of various clinical disorders. These cells have a great capacity for proliferation and differentiation, do not form teratomas when transplanted into animal models and their stemness seems to be between embryonic cells and adult mesenchymal cells. Prior to their use in cell therapy they must be cultured and expanded in vitro, but the effect this process has on their fitness, a determining factor for the success or failure of cell therapy, is unknown. We undertook a follow-up of gene and microRNAs (miRNAs) expression using microarray of a hAMSC population kept under in vitro culture conditions for the first 15 passages. Significant changes were noted in the expression of various mRNAs and miRNAs, particularly down-regulation of TP53, increased expression of hsa-miR-125a and up-regulation of CDKN2D. The variations in TP53 and hsa-miR-125a may act as an indicator of the stemness of the hAMSC, whereas CDKN2D may indicate the reduction in the proliferative capacity of these cells in a TP53-independent mechanism. The genes described in this study will help evaluate the fitness of hAMSC, thus guaranteeing their biological quality for use in regenerative medicine.

ORGANISM(S): Mus musculus Rattus norvegicus synthetic construct Homo sapiens

PROVIDER: GSE30065 | GEO | 2011/06/18

SECONDARY ACCESSION(S): PRJNA155155

REPOSITORIES: GEO

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Cultured human amniotic fluid-derived mesenchymal stromal cells [miRXplore data]

Project description:Human amniotic fluid-derived mesenchymal stromal cells (hAMSC) have become one of the main cell populations used in regenerative medicine and for the study of various clinical disorders. These cells have a great capacity for proliferation and differentiation, do not form teratomas when transplanted into animal models and their stemness seems to be between embryonic cells and adult mesenchymal cells. Prior to their use in cell therapy they must be cultured and expanded in vitro, but the effect this process has on their fitness, a determining factor for the success or failure of cell therapy, is unknown. We undertook a follow-up of gene and microRNAs (miRNAs) expression using microarray of a hAMSC population kept under in vitro culture conditions for the first 15 passages. Significant changes were noted in the expression of various mRNAs and miRNAs, particularly down-regulation of TP53, increased expression of hsa-miR-125a and up-regulation of CDKN2D. The variations in TP53 and hsa-miR-125a may act as an indicator of the stemness of the hAMSC, whereas CDKN2D may indicate the reduction in the proliferative capacity of these cells in a TP53-independent mechanism. The genes described in this study will help evaluate the fitness of hAMSC, thus guaranteeing their biological quality for use in regenerative medicine. miRNA patterns were studied in in vitro culture samples of human amniotic fluid-derived mesenchymal stromal cells at passages P1, P3, P5 and P10 with miRXplorer microarrays using a synthetic pool of miRNAs. Cell pellets of 5 x 10E5 cells were cryopreserved in liquid nitrogen and sent to Miltenyi Biotec for their analysis.

2011-06-18 | E-GEOD-30065 | biostudies-arrayexpress

Cultured human amniotic fluid-derived mesenchymal stromal cells [PIQOR data]

2011-06-18 | GSE30064 | GEO

Cultured human amniotic fluid-derived mesenchymal stromal cells [PIQOR data]

Project description:Human amniotic fluid-derived mesenchymal stromal cells (hAMSC) have become one of the main cell populations used in regenerative medicine and for the study of various clinical disorders. These cells have a great capacity for proliferation and differentiation, do not form teratomas when transplanted into animal models and their stemness seems to be between embryonic cells and adult mesenchymal cells. Prior to their use in cell therapy they must be cultured and expanded in vitro, but the effect this process has on their fitness, a determining factor for the success or failure of cell therapy, is unknown. We undertook a follow-up of gene and microRNAs (miRNAs) expression using microarray of a hAMSC population kept under in vitro culture conditions for the first 15 passages. Significant changes were noted in the expression of various mRNAs and miRNAs, particularly down-regulation of TP53, increased expression of hsa-miR-125a and up-regulation of CDKN2D. The variations in TP53 and hsa-miR-125a may act as an indicator of the stemness of the hAMSC, whereas CDKN2D may indicate the reduction in the proliferative capacity of these cells in a TP53-independent mechanism. The genes described in this study will help evaluate the fitness of hAMSC, thus guaranteeing their biological quality for use in regenerative medicine. The variations in the expression patterns of the mRNAs were studied using topic-defined PIQOR Stem Cell microarrays. Human amniotic fluid-derived mesenchymal stromal cells pellets of 5 x 10E5 cells were cryopreserved in liquid nitrogen and send to Miltenyi Biotec for their analysis. A pool of non-stimulated lymphocytes (non-mitotic cell cycle) obtained by Ficoll isolation in the Immunology Service of Carlos Haya hospital was used as a control of the expression.

2011-06-18 | E-GEOD-30064 | biostudies-arrayexpress

Expression data on Treg cells isolated from wildtype mice and miR-125a deficient mice

2015-05-08 | GSE66608 | GEO

RNA-Seq of inflammatory stimulated HUVECs transfected with microRNA-125a-5p or negative control.

Project description:Purpose: In recent years, research has revealed the role of microRNAs (miRs) as important regulators of endothelial function. Notably, miR-125a is upregulated in the blood of patients with acute vascular diseases. Since miRNAs act out their function in networks, we aimed at investigating the presence of a miR-125a-related network regulating the endothelial barrier. Method: We investigated transcriptional changes of human umbilical cord endothelial cells (HUVEC) after miR-125a overexpression in an acute inflammatory in-vitro setting using Next Generation Sequencing. Briefly, primary HUVECs from five different donors were transfected either with hsa-miR-125a-5p or negative control (NC). After transfection HUVECs were cultivated for 18 hours and stimulated with TNF (25 ng/ml) for additional 6 hours. We compared the following groups: NC (18047-0001, 18047-0002, 18047-0003, 18047-0004, 18047-0005) versus miR125a: (18047-0006, 18047-0007, 18047-0008, 18047-0009, 18047-0010) Results: MiR-125a overexpression in inflammatory activated HUVECs significantly altered gene expression of 468 genes (FC: +1.25 to -1.25 p<0.01). We found eight potential miR-125a direct target genes involved in endothelial barrier function.

2022-02-07 | GSE196161 | GEO

Multicellular Transcriptional Analysis of Mammalian Heart Regeneration

Project description:The inability of the adult mammalian heart to regenerate following injury represents a major barrier in cardiovascular medicine. In contrast, the neonatal mammalian heart retains a transient capacity for regeneration, which is lost shortly after birth. Defining the molecular mechanisms that govern regenerative capacity in the neonatal period remains a central goal in cardiac biology. Here, we construct a transcriptional atlas of multiple cardiac cell populations, which enables comparative analyses of the regenerative (neonatal) versus non-regenerative (adult) state for the first time. This work provides a comprehensive transcriptional resource of multiple cardiac cell populations during cardiac development, repair and regeneration. Our findings define a transcriptional program underpinning the neonatal regenerative state and identifies an epigenetic barrier to re-induction of the regenerative program in adult cardiomyocytes.

2017-07-24 | GSE95755 | GEO

Unravelling Heterogeneity of Amplified Human Amniotic Fluid Stem Cells Sub-Populations

Project description:Human amniotic fluid stem cells (hAFSCs) are broadly multipotent immature progenitor cells with high self-renewal and no tumorigenic properties. These cells, even amplified, present very variable morphology, density, intracellular composition and stemness potential, and this heterogeneity can hinder their characterization and potential use in regenerative medicine. Celector® (Stem Sel ltd.) is a new technology that exploits the Non-Equilibrium Earth Gravity Assisted Field Flow Fractionation principles to characterize and label-free sort stem cells based on their solely physical characteristics without any manipulation. Viable cells are collected and used for further studies or direct applications. In order to understand the intrapopulation heterogeneity, various fractions of hAFSCs were isolated using the Celector® profile and live imaging feature. The gene expression profile of each fraction was analysed using whole-transcriptome sequencing (RNAseq). Gene Set Enrichment Analysis identified significant differential expression in pathways related to Stemness, DNA repair, E2F targets, G2M checkpoint, hypoxia, EM transition, mTORC1 signalling, Unfold Protein Response and p53 signalling. These differences were validated by RT-PCR, immunofluorescence and differentiation assays. Interestingly, the different fractions showed distinct and unique stemness properties. 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 certain cellular fractions with the highest potential to use in regenerative medicine.

2021-01-13 | GSE164692 | GEO

Expression data on Treg cells isolated from wildtype mice and miR-125a deficient mice

Project description:We found miR-125a was a key regulator that stabilizes the commitment and immunoregulatory capacity of Treg cells.To gain insights into the general functional features of miR-125a-deficient Treg cells, we performed a genome-wide gene array analysis on Treg population isolated from the spleens of 6 to 8-week-old miR-125a-deficient and WT mice We sorted CD4+CD25hi Treg population from the spleens of 6 to 8-week-old miR-125a-deficient and their littermate WT mice. Cells were collected and total RNA was extracted for Affymetrix GeneChip®Mouse Genome 430 2.0 Array

2015-05-08 | E-GEOD-66608 | biostudies-arrayexpress

Regeneration in the pituitary after cell-ablation injury: time-related aspects and molecular analysis

Project description:We recently showed that the mouse pituitary holds regenerative competence. Young-adult GHCre/iDTR mice, expressing diphtheria toxin (DT)-receptor in growth hormone (GH)-producing cells, regenerate the cells ablated by 3-day DT treatment (3DT) up to 60% after 5 months. The pituitary's stem cells were found to participate in this restoration process. Here, we characterized this regenerative capacity in relation to age and recovery period, and started to search for underlying molecular mechanisms. Extending the recovery period (to 19 months) does not result in higher regeneration levels. In addition, the regenerative competence disappears at ageing, coinciding with a reduction in pituitary stem cell number and fitness. Surprisingly, prolonging the DT treatment period in young-adult mice to 10 days (10DT) completely blocks the regeneration. The stem cell compartment still reacts by promptly expanding, and retains in vitro stem cell functionality. To obtain a first view on molecular grounds underlying reparative capacity and/or failure, the stem cell-clustering side population (SC-SP) was analyzed by whole-genome expression analysis. A number of stemness factors and components of the Notch, Shh, Wnt, epithelial-mesenchymal transition (EMT) and Hippo pathways are higher expressed in the SC-SP of the regenerating pituitary (after 3DT) when compared to the basal gland and to the non-regenerating pituitary (after 10DT). Taken together, the regenerative capacity of the pituitary is limited both in age-related terms and final efficacy, and appears to rely on stem cell-associated pathway activation. Dissection of the molecular profiles may eventually identify targets to induce or boost regeneration in situations of (injury-related) pituitary deficiency.

2018-09-03 | GSE72992 | GEO

Extracellular-acidosis restricts one-carbon metabolism and preserves T cell stemness

Project description:The accumulation of acidic metabolic waste products within the tumor microenvironment profoundly inhibits effector functions of tumor-infiltrating lymphocytes (TILs). However, it is unclear whether extracellular-acidosis impacts T cell metabolic fitness and differentiation status. Here, we surprisingly found that prolonged acid exposure reprogrammed T cell intracellular metabolism and mitochondrial fitness, and preserved T cell stemness. Elevated extracellular-acidosis impaired methionine uptake and metabolism via downregulating SLC7A5, therefore altering H3K27me3 deposition at the promoters of crucial T cell stemness genes. These changes promoted the maintenance of a “stem-like memory” state and improved long-term in vivo persistence and anti-tumor efficacy. Our findings not only reveal the unexpected roles of extracellular-acidosis in the maintenance of stem-like properties of T cells, but also advance our understanding of the direct involvement of methionine metabolism in T cell stemness.

2023-01-17 | MTBLS6661 | MetaboLights

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