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Series GSE54574 Query DataSets for GSE54574
Status Public on Mar 07, 2016
Title Direct Conversion from Mouse Fibroblasts Informs the Identification of Hemogenic Precursor Cells In Vivo
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Definitive hematopoiesis emerges via an endothelial-to-hematopoietic transition in the aorta-gonad-mesonephros (AGM) region and placenta. We have recently demonstrated the induction of hematopoietic stem/progenitors (HSPCs) from mouse fibroblasts with a combination of transcription factors progressing through endothelial-like precursors. Here, guided by our in vitro programming experiments we analyzed mouse placentas for the presence of the precursor phenotype. We identified a small population of CD34+ Sca1+Prom1+ (34PS) cells in mid-gestation placentas that do not express the pan-hematopoietic marker CD45. After isolation and culture 34PS cells acquire CD45 and generate large hematopoietic as well as cobblestone colonies. Prom1+ cells localize to the placental vascular labyrinth where HSPCs emerge. 34PS cells express markers associated with the hemogenic endothelium (CD31, Tie2, VE-Cadherin, Sox17, Runx1, Scl) and also markers identified by direct induction (Itga6/CD49f). This population is heterogeneous for the early hematopoietic marker CD41 and expresses the programming transcription factors. Remarkably, global gene expression profiles of placental 34PS cells correlate with AGM-derived hemogenic endothelium and fibroblast-derived precursors. Finally, when co-cultured with stroma placental 34PS cells give rise to B/T lymphoid cells as well as mixed colonies containing erythroid, myeloid and megakaryocytic cell lineages. In summary, we show that direct in vitro conversion provided a cell surface phenotype for the isolation of hemogenic precursors in vivo. Our findings provide insights into the specification of definitive hemogenesis in the placenta, in depth characterization of hemogenic precursor populations and the first evidence that direct in vitro conversion approaches can be used as a valuable tool to address basic developmental questions in vivo.
 
Overall design mRNAseq profiling on populations isolated by selected marker fluorescence activated cell sorting
The 'E10_E12_HSPC_SingleCell_FPKM.txt.gz' contains the processed data for GSM1890353-GSM1890496.
 
Contributor(s) Pereira C, Moore K, Lemischka IR
Citation(s) 26954547
Submission date Jan 30, 2014
Last update date May 15, 2019
Contact name Betty Chang
E-mail(s) filipe.pereira@uc-biotech.pt
Organization name Mount Sinai School of Medicine
Street address 1425 Madison Ave
City New York
State/province NY
ZIP/Postal code 10029
Country USA
 
Platforms (1)
GPL13112 Illumina HiSeq 2000 (Mus musculus)
Samples (167)
GSM1319198 E10_cKit-_2
GSM1319199 E10_cKit-_3
GSM1319200 E10_cKit+_1
Relations
BioProject PRJNA237041
SRA SRP036078

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE54574_E10_E12_HSPC_SingleCell_FPKM.txt.gz 3.8 Mb (ftp)(http) TXT
GSE54574_Placenta_FPKM_table.txt.gz 1.1 Mb (ftp)(http) TXT
GSE54574_Ter119_FPKM.txt.gz 459.6 Kb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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