Proteasome Function is Required for Platelet Production
ABSTRACT: RNA-seq expression analysis of transcripts encoding proteasome subunits in human CD34+ cord blood cell-derived megakaryocytes and mouse bone marrow-derived megakaryocytes. Analysis of transcript expression in human CD34+ cord blood cell-derived megakaryocytes and mouse bone marrow-derived megakaryocytes.
Project description:A human bone marrow-derived mesenchymal stromal cell (MSCs) and cord blood-derived CD34+ stem cell co-culture system was set up in order to evaluate the proliferative and differentiative effects induced by MSCs on CD34+ stem cells, and the reciprocal influences on gene expression profiles Overall design: Bone marrow-derived mesenchymal stromal cells (BM-MSCs) were seeded on a plastic support. Upon reaching 80% of confluence, previously isolated CD34+ cord blood-derived hematopoietic stem cells (CB-CD34+) were plated onto a MSCs layer. After 10 days of culture, cells grown in suspension were collected in plastic tubes, while adherent cells were harvested by trypsinization. Two different fractions of HSCs were obtained: the SN-fraction, representing the HSCs grown in the supernatant of the co-cultures, and the AD-fraction, representing the HSCs grown directly in contact with the stromal layer (co-coltured MSCs). Single cultures of BM-MSCs and CB-CD34+ cells served as controls, namely MSCs-alone and CB-alone
Project description:Global gene expression analysis of (a) human embryonic stem cells, (b) adult fibroblasts with and without nucleofection of SOKM, (c) CD34+ cord blood cells at various time points during induction of pluripotency with SOKM, with or without co-culture with bone marrow stromal cells (BMSC), and (d) resulting stromal primed and non-stromal primed cord blood CD34+ myeloid iPSC Overall design: Total RNA was harvested from adult fibroblasts, adult fibroblasts three days after nucleofection with 4F (SOKM), human myeloid progenitors: Non-nucloefected CD34+ Day -3 unstimulated cord blood (CB) cells, Day 0 growth factor stimulated CB cells, Day +3 CB cells with or without nucleofection at Day zero and with or without Day zero to Day +3 co-culture with bone marrow stromal cells (BMSC), followed by FACS-purification of CD45+ myeloid progenitors, Day 23 CB cells nucleofected on day zero and co-cultured on BMSC from Day Zero to Day +3, undifferentiated H9 human embryonic stem cells, and low passage episomal stromal-primed iPSC derived from growth-factor activated CD34+ myeloid progenitors from cord blood or GCSF mobilized peripheral blood. Illumina HumanHT-12 V4.0 expression beadchips were used for all samples in this series. Three independent samples of each condition were each run on individual microarrays.
Project description:Gene expression profiles of CD34+CD38- stem cells and more differentiated CD34+CD38+ progenitor cells were compared. Comparison of expression profiles of hematopoietic stem cells from fetal liver, umbilical cord blood, bone marrow and mobilized pheripheral blood allowed us to identify a unique set of genes with conserved expression during ontogeny. Experiment Overall Design: CD34+CD38- en CD34+CD38+ cell populations were isolated by cell sorting from human Bone Marrow, mobilized peripheral blood, umbilical cord blood and fetal liver. Total RNA was isolated from each cell population followed by the synthesis of biotinylated cRNA. After fragmentation the biotinylated cRNA was hybridized to affymetrix U133A chips.
Project description:Transcription profiling analysis was performed on purified CD34+ cell lines (Cord Blood CD34+) treated with ExtracellularVescicles (EVs) isolated from bone marrow mesenchymal stem cells (BM-MSC).
Project description:Microarray gene expression data is used to compare the transcriptomics of hiPSC, hiPSC derived forward programmed-, hiPSC derived directed differentiated- megakaryoctes AND cord- blood derived megakaryocytes Overall design: Total RNA obtained from human induced plutipotent stem cells (hiPSC), megakaryocytes differentiated in vitro from haematopoetic stem cells obtained from cord blood and megakaryocytes differentiated from hiPSC
Project description:An increased number of circulating CD34+ hematopoietic progenitors (HP) and a prominent amplification of dystrophic megakaryocytes (MK) are observed in PMF patients. As transcriptome data from CD34+ hematopoietic progenitors showed modulations of FLT3 and MAP kinase expression independently of the JAK2V617F mutation status Overall design: Transcriptome analysis was performed on circulating CD34+ cells from PMF patients using Agilent 22K microarray and compared to CD34+ cells from blood and bone marrow from un-mobilized healthy donors. Indirect map: each tested sample was hybridzed with reference probe
Project description:An increased number of circulating CD34+ hematopoietic progenitors (HP) and a prominent amplification of dystrophic megakaryocytes (MK) are observed in PMF patients. As transcriptome data from CD34+ hematopoietic progenitors showed modulations of FLT3 and MAP kinase expression independently of the JAK2V617F mutation status Transcriptome analysis was performed on circulating CD34+ cells from PMF patients using Agilent 22K microarray and compared to CD34+ cells from blood and bone marrow from un-mobilized healthy donors. Indirect map: each tested sample was hybridzed with reference probe
Project description:Investigation of human hematopoietic stem cells gene expression patterns originating from different stages of ontogeny including fetal blood, cord blood, bone marrow, and mobilized peripheral blood in Lin-CD34+CD38- versus Lin-CD34+CD38+ populations. Experiment Overall Design: this experiment include 7 samples and 42 replicates
Project description:In order to identify genes associated with the engraftment potential of human hematopoietic stem cells, we have employed whole genome microarray expression profiling of G0 and G1 phase CD34+ cells derived from bone marrow, mobilized peripheral blood, and umbilical cord blood. Samples were collected from healthy adult volunteers after obtaining informed consent according to the guidelines of the Investigational Review Board of Indiana University School of Medicine. CD34+ cells were selected and fractionated into G0 and G1 phases of cell cycle on a flow cytometer. Purity of sorted cells was further confirmed by qRT-PCR by measuring the relative expression of Ki67. Sorted cells were subjeccted to microarray analysis. Overall design: Three biological replicates of sorted and confirmed G0 and G1 cells from bone marrow, mobilized peripheral blood, and umbilical cord blood (total of eighteen samples) were subjected to microarray analysis. To generate distinct and unique sets of data, we did not pool multiple samples from any tissue studied so that each sample or its replicate was from a single donor.