The human reticulocyte transcriptome (HG-U133_Plus2.0)
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE17639 | GEO | 2009/12/01
SECONDARY ACCESSION(S): PRJNA118581
REPOSITORIES: GEO
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Project description:RNA from circulating blood reticulocytes was utilized to provide a robust description of genes transcribed at the final stages of erythroblast maturation. After depletion of leukocytes and platelets, Affymetrix HG-U133Plus 2.0 arrays were hybridized with probe from total RNA isolated from blood sampled from 6 umbilical cords and 6 healthy adult humans. Peripheral blood from 6 normal adults and 6 human umbilical cords were purified to obtain packed red blood cells and their purity was assessed by CELL-DYN4000. The contaminated WBC count was less than 1 out of 1 million cells. Total RNA from purified reticulocytes included in the purified red blood cells was extracted, labeled, and hybridized onto Affymetrix HG-U133Plus 2.0 arrays.
2009-11-30 | E-GEOD-17639 | biostudies-arrayexpress
Project description:RNA from circulating blood reticulocytes was utilized to provide a robust description of genes transcribed at the final stages of erythroblast maturation. After depletion of leukocytes and platelets, Affymetrix HG-U133 arrays were hybridized with probe from total RNA isolated from blood sampled from 14 umbilical cords and 14 healthy adult humans. Keywords: Adult vs. fetal reticulocyte transcriptome comparison
2007-03-26 | GSE6236 | GEO
Project description:RNA from circulating blood reticulocytes was utilized to provide a robust description of genes transcribed at the final stages of erythroblast maturation. After depletion of leukocytes and platelets, Affymetrix HG-U133 arrays were hybridized with probe from total RNA isolated from blood sampled from 14 umbilical cords and 14 healthy adult humans. Experiment Overall Design: Peripheral blood from 14 normal adult and 14 human umbilical cord were purified to obtain packed red blood cells and their purity was assessed by CELL-DYN4000. The contaminated WBC count was less than 1 out of 1 million cells. Total RNA from purified reticulocytes included in the purified red blood cells was extracted, labeled, and hybridized onto Affymetrix HG-U133 A and B arrays.
2007-11-01 | E-GEOD-6236 | biostudies-arrayexpress
Project description:Reticulocytes were purified from fresh blood samples obtained from 10 healthy adult volunteers (5 women and 5 men) after informed consent. The donors had normal blood cell counts, blood smears, hemoglobin electrophoresis, red cells membrane resistance tests and no biological evidence of hemolysis. Whole blood samples were centrifuged, the supernatant and the buffy coat containing white blood cells (WBC) and platelets were removed. The red cells pellet was then purified using the method described by Brun et al. Purity, assessed by Hematology Flow Cytometer was 1 leukocyte per 15 millions RBC and efficiency of purification process was 99.8% and 1 platelet per 10 000 RBC. Keywords: other
2004-10-23 | GSE1866 | GEO
Project description:Mouse models have proven invaluable for understanding erythropoiesis. Here, we describe an autosomal recessive inherited anemia in the mouse mutant hem6. Hematologic and transplantation analyses revealed a mild, congenital, hypochromic, microcytic anemia intrinsic to the hematopoietic system that is associated with a decreased red blood cell zinc protoporphyrin to heme ratio, indicative of porphyrin insufficiency. Iron uptake experiments showed that hem6 reticulocytes are defective in heme production, but not cellular iron uptake defects. Male hem6 mice are infertile due to defects in sperm structure and motility. Through positional cloning and BAC complementation, we identified the gene responsible for the hem6 anemia. We hypothesized that the relative deficiency in erythroid-specific mRNAs in hem6 reticulocytes might be due to decreased mRNA stability. Indeed, serial microarray analysis of reticulocytes aged in vitro showed that numerous, abundantly expressed erythroid-specific transcripts decayed at faster rates in hem6 reticulocytes compared to control reticulocytes. Furthermore, these mRNAs also have progressively shorter poly (A) tails, suggesting a mechanism for the increased rate of decay. Keywords: serial time points Reticulocyte rich blood were collected and cultured ex vivo for 24 hours, samples were collected at 0, 12,24 hours for microarray analysis. There are 3 wild type (wt) biological replicates and 5 mutant (mut) biological replicates in each time point.
2009-10-12 | E-GEOD-16174 | biostudies-arrayexpress
Project description:Mouse models have proven invaluable for understanding erythropoiesis. Here, we describe an autosomal recessive inherited anemia in the mouse mutant hem6. Hematologic and transplantation analyses revealed a mild, congenital, hypochromic, microcytic anemia intrinsic to the hematopoietic system that is associated with a decreased red blood cell zinc protoporphyrin to heme ratio, indicative of porphyrin insufficiency. Iron uptake experiments showed that hem6 reticulocytes are defective in heme production, but not cellular iron uptake defects. Male hem6 mice are infertile due to defects in sperm structure and motility. Through positional cloning and BAC complementation, we identified the gene responsible for the hem6 anemia. We hypothesized that the relative deficiency in erythroid-specific mRNAs in hem6 reticulocytes might be due to decreased mRNA stability. Indeed, serial microarray analysis of reticulocytes aged in vitro showed that numerous, abundantly expressed erythroid-specific transcripts decayed at faster rates in hem6 reticulocytes compared to control reticulocytes. Furthermore, these mRNAs also have progressively shorter poly (A) tails, suggesting a mechanism for the increased rate of decay. Keywords: serial time points
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Project description:mRNA from 59 primary colorectal tumour samples were extracted and hybridized to HG-U133Plus 2.0 expression arrays. Mutation status for several genes were determined using Sanger sequencing.
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Project description:Human reticulocytes were enriched from human umbilical blood, and cultured in vitro for the production of reticulocyte-derived exosomes (HuRex)to unveil their proteomic composition. As research on the molecular-cargo of exosomes is largely confounded by the lack of a “gold-standard” methodology, we applied several approaches for obtaining HuRex prior to MS. HuRex preparations were obtained from cultures in absence of serum (AS) from three donors and isolated by means of size-exclusion chromatography (SEC) (n=3) and ultracentrifugation (UC) (n=3). HuRex preparations in the presence of serum were obtained from three other donors and purified by SEC (n=3) and UC (n=1).
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Project description:Regular blood transfusion is the cornerstone of care for patients with red blood cell (RBC) disorders such as thalassemia or sickle cell disease. With repeated transfusion, alloimmunisation often occurs due to incompatibility at the level of minor blood group antigens. We use CRISPR-mediated genome editing of an immortalised human erythroblast cell line (BEL-A) to generate multiple enucleation competent cell lines deficient in individual blood groups. Edits are combined to generate a single cell line deficient in multiple antigens responsible for the most common transfusion incompatibilities: ABO (Bombay phenotype), Rh (Rhnull), Kell (K0), Duffy (Duffynull), GPB (S- s- U-). These cells can be differentiated to generate deformable reticulocytes, illustrating the capacity for coexistence of multiple rare blood group antigen null phenotypes. This study provides the first proof-of-principle demonstration of combinatorial CRISPR-mediated blood group gene editing to generate customisable or multi-compatible RBCs for diagnostic reagents or recipients with complicated matching requirements.
2018-04-26 | PXD009291 | Pride