Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Microarray gene expression analysis of mouse fetal liver erythroblasts after plek2 knockdown

ABSTRACT: The goal of this experiment is to determine the expression levels of the genes that are influenced by the downregulation of plek2. TER119 negative mouse fetal liver erythroblasts were purified and infected with control and plek2 shRNA retroviruses and cultured in Epo free medium for 24 hours followed by Epo medium for 24 hours. Microarray gene expression profile analysis was performed using an Illumina platform with a biological triplicate of the same experiment.

ORGANISM(S): Mus musculus

SUBMITTER: Peng Ji

PROVIDER: E-GEOD-48332 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The supply of red blood cells (RBCs) is not sufficient in many developing countries or in developed countries for patients who need chronic transfusion from best-matched donors. Ex vivo expansion and maturation of human erythroid precursor cells (erythroblasts) could represent a potential solution. Proliferating erythroblasts can be expanded from human umbilical cord blood mononuclear cells (CB MNCs) ex vivo for 10^6-10^7 fold (in ~50 days) before undergoing senescence. Here, we report that ectopic expression of three to four genetic factors that have been used for iPS cell derivation enables CB-derived erythroblasts to undergo extended ex vivo expansion (M-bM-^IM-%10^51 fold in ~9 months) in a defined suspension culture condition without change of cell identity or function. These vastly expanding erythroblasts maintain homogeneously immature erythroblast phenotypes, a normal diploid karyotype and dependence on specific combination of cytokines and hormone for survival and proliferation throughout the continuous expansion period. When switched to a culture condition for terminal maturation, these immortalized erythroblasts gradually exit cell cycle, decrease cell size, accumulate hemoglobin, condense nuclei and eventually give rise to enucleated hemoglobin-containing erythrocytes. Our result may ultimately lead to the development of unlimited sources of cultured RBCs for optimally-matched or personalized transfusion medicine. We compared the global gene expression profiles of different human cell types: iE: immortalized erythroblasts generated by genetic reprogramming from pCBE; pCBE: primary cord blood-derived erythroblasts; CD34+: CD34+ purified hematopoietic stem/progenitor cells from adult blood or fetal liver; TF-1: a human erythroleukemia cell line; ESC: human embryonic stem cells; iPSCs: human induced pluripotent stem cells. We want to see the relationship among these cell types. We included multiple samples (biological replicates) for most cell types.

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Microarray gene expression analysis of mouse fetal liver erythroblasts after plek2 knockdown

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