Project description:Normal healthy cells (monocytes, promyelocytes, polymorphonuclear cells, B cells, T cells, CD34+CD38- HSPCs) run as controls for TCGA AML marker publication. Bone marrow cells collected from healthy donors were sorted and DNA extracted at Washington University in St. Louis, microarrays were then run at USC
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.
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. 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.
Project description:CD34+ progentitors were isolated from the bone marrow of three healthy volunteers. CD34+CD71+CD45RA- were FACS sorted to enrich for erythroid progenitors. The cells were cultured for four hours with or without EPO in combination with LY294002, and harvested for RNA extraction, amplification and expression analysis.
Project description:miRNA expression in a patient with AML comparing with pooled CD34 hematopoietic progenitor cells from 5 healthy volunteers RNA from bone marrow of a patient with AML with more than 90% blast and RNA pooled from 5 volunteers with CD34+ cells selected by automacs from bone marrow
