biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsSatoshi Kondotranscription profiling by arrayMus musculusMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-GEOD-46158Self-renewal potential and multipotency are hallmarks of the stem cell1-3. It is generally conceived that acquisition of such a stemness requires rejuvenation of somatic cells through reprogramming of genetic and epigenetic status. We show here that a simple blockage of cell differentiation is sufficient to make stem cells. By over-expression of transcriptional repressor Id3 in murine hematopoitic progenitor cells and cultivation of these cells in B cell induction conditions, cells exhibit developmental arrest and enter a self-renewal cycle. These cells can be maintained in vitro almost limitlessly, and the long-term cultured cells exhibited robust multi-lineage reconstitution when transferred into irradiated mice. These cells can be cloned and re-expanded with 50% of plating efficiency, ibiostudies-arrayexpressGrowth Protocol - RPMI 1640 medium supplemented with 10% FCS, L-glutamine (2 mM), sodium pyruvate (1 mM), sodium bicarbonate (2 mg/ml), nonessential amino acid solution (0.1 mM), 2-ME (5 x 10-5 M), streptomycin (100 mg/ml), and penicillin (100 U/ml).Labeling - Amino Allyl aRNA was synthesis by Amino Allyl MessageAmp II aRNA Amplification Kit (Ambion#1753). CyeDye Coupling and fragmentation were performed as the protocol supplied by TORAY Industries, Inc..Hybridization - Hybridized for 16 h at 37 C with rotary shake (250 rpm). Hybridization buffer and washing protocol was followed by the protocol supplied by TORAY Industries, Inc..Nucleic Acid Extraction - Total RNA from each sample (1x10^5 - 1x10~6 cells) was purified by RNeasy mini Kit (Qiagen)Scaning - ScanArray Express (PerkinElmer Japan Co., Ltd.) was used for scanning. Images were quantified using GenePix Pro version 6.0(Axon Instruments).MIAME ScoreRaw DataOrganizationAssays and DataProcessed DataMAGE-TAB FilesArray DesignsTomokatsu IkawaSatoshi KondoHiroshi KawamotoData Transformation - The raw data of each spot was normalized by substitution with a mean intensity of the background signal determined by all blank spots’ signal intensities of 95% confidence intervals. Measurements of both duplicate spots with the signal intensities greater than 2 standard deviations (SD) of the background signal intensity were considered to be valid. A relative expression level of a given miRNA was calculated by comparing the signal intensities of the averaged valid spots with their mean value throughout the microarray experiments. ID_REF = VALUE = Normalized signal intensityfalseMicroarray analysis of IdHP cells compared to LKS, Control ProB and E2A-/- progenitorsSelf-renewal potential and multipotency are hallmarks of the stem cell1-3. It is generally conceived that acquisition of such a stemness requires rejuvenation of somatic cells through reprogramming of genetic and epigenetic status. We show here that a simple blockage of cell differentiation is sufficient to make stem cells. By over-expression of transcriptional repressor Id3 in murine hematopoitic progenitor cells and cultivation of these cells in B cell induction conditions, cells exhibit developmental arrest and enter a self-renewal cycle. These cells can be maintained in vitro almost limitlessly, and the long-term cultured cells exhibited robust multi-lineage reconstitution when transferred into irradiated mice. These cells can be cloned and re-expanded with 50% of plating efficiency, i2014-06-01T00:00:00Z2023-08-27T14:58:58.285Z2021-10-01T18:01:52ZE-GEOD-46158GSE46158EFO_0002768