Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Comparison of Akt activated cells and placental mesenchymal progenitors

ABSTRACT: The goal of our experiment is to compare the cytokines and membrane factors that are differentially expressed between akt-activated endothelial cells and mesenchymal progenitors. Those two cell type have previously been described as potential niche for HSC expansion in vivo and in vitro. We isolated placental mesenchymal progenitors as previously described (Raynaud CM. et al. 2012 stem cell international). HUVECs cells were also isolated from cord and transfected with E4ORF1 as previously described (Seandel M. et al. 2008 PNAS). Both cell type were grown separately 3 days in X-Vivo media complemented with TPO, Flt3 and SCF (HSc amplification media). Pl-MPs and E4+ECs RNA was isolated using Trizol reagent followed by additional purification using RNAeasy extraction kit from Qiagen (74106, QIAGEN) with RNA yields that produces satisfactory microarray data. Two quality control measures were carried out: (1) A spectrophotometric analysis (2) A size fractionation procedure using a microfluidics instrument (Agilent Technologies). 200 ng of total RNA were analyzed on Affymetrix GeneChip Human Genome U133 Plus 2.0 Array. Data were analyzed using Parteck Software (V6.09.1110-6; Affimetrix).

ORGANISM(S): Homo sapiens

SUBMITTER: Christophe Raynaud

PROVIDER: E-MEXP-3725 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Endothelial cells provide a niche for placental hematopoietic stem/progenitor cell expansion through broad transcriptomic modification.

Raynaud Christophe M CM Butler Jason M JM Halabi Najeeb M NM Ahmad Faizzan S FS Ahmed Badereldeen B Rafii Shahin S Rafii Arash A

Stem cell research 20130809 3

Umbilical cord blood (UCB) is an attractive source of hematopoietic stem cells (HSCs). However, the number of HSCs in UCB is limited, and attempts to amplify them in vitro remain inefficient. Several publications have documented amplification of hematopoietic stem/progenitor cells (HSPCs) on endothelial or mesenchymal cells, but the lack of homogeneity in culture conditions and HSC definition impairs direct comparison of these results. We investigated the ability of different feeder layers, mese ...[more]

PMID: 23978474

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Project description:Hematopoietic stem cells (HSCs) primarily reside in the bone marrow where signals generated by stromal cells regulate their self-renewal, proliferation, and trafficking. Endosteal osteoblasts and perivascular stromal cells including endothelial cells3, CXCL12-abundant reticular (CAR) cells, leptin-receptor positive stromal cells, and nestin-GFP positive mesenchymal progenitors have all been implicated in HSC maintenance. However, it is unclear if specific hematopoietic progenitor cell (HPC) subsets reside in distinct niches defined by the surrounding stromal cells and the regulatory molecules they produce. CXCL12 (stromal-derived factor-1, SDF-1) regulates both HSCs and lymphoid progenitors and is expressed by all of these stromal cell populations. Here, we selectively deleted Cxcl12 from candidate niche stromal cell populations and characterized the effect on HPCs. Deletion of Cxcl12 from mineralizing osteoblasts has no effect on HSCs or lymphoid progenitors. Deletion of Cxcl12 from osterix-expressing stromal cells, which includes CAR cells and osteoblasts, results in constitutive HPC mobilization and a loss of B lymphoid progenitors, but HSC function is normal. Cxcl12 deletion in endothelial cells results in a modest loss of long-term repopulating activity. Strikingly, deletion of Cxcl12 in nestin-negative mesenchymal progenitors using Prx1-Cre is associated with a marked loss of HSCs, long-term repopulating activity, HSC quiescence, and common lymphoid progenitors. These data suggest that osterix-expressing stromal cells comprise a distinct niche that supports B lymphoid progenitors and retains HPC in the bone marrow, while expression of CXCL12 from stromal cells in the perivascular region, including endothelial cells and mesenchymal progenitors, support HSCs. Total of three samples of two groups analyzed. Replica samples of CXCL12-abundant reticular (CAR) cells from two CXCL12-GFP knock-in mice and a combined sample of PDGFRa+ Sca+ CD45- lineage- cells from three Prx1-Cre Rosa26Ai9/+ Cxcl12gfp/+ mice were used and analyzed.

Dataset Information

Comparison of Akt activated cells and placental mesenchymal progenitors

Publications

Endothelial cells provide a niche for placental hematopoietic stem/progenitor cell expansion through broad transcriptomic modification.

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