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RNA-sequencing of human peripheral blood mononuclear cells (PB MNCs) and induced neural progenitor cells derived from PB MNCs

ABSTRACT: RNA-sequencing of human peripheral blood mononuclear cells (PB MNCs) and induced neural progenitor cells derived from PB MNCs

PROVIDER: PRJNA421444 | ENA |

REPOSITORIES: ENA

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RNA-sequencing of human peripheral blood mononuclear cells (PB MNCs) and induced neural progenitor cells derived from PB MNCs

Project description:We have successfully generated human induced neural progenitor cells (iNPCs) from peripheral blood mononuclear cells (PB MNCs). To gain further insights into the transcriptional activation and cellular identity of human iNPCs, we compared the transcriptome of human iNPCs with PB MNCs and published database CORTECON and BrainSpan.

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Project description:The use of induced pluripotent stem cells (iPSCs) is an exciting frontier in the study and treatment of human diseases through the generation of specific cell types. Here we show the derivation of iPSCs from human non-mobilized peripheral blood (PB) and bone marrow (BM) mononuclear cells (MNCs) by retroviral transduction of OCT3/4, SOX2, KLF4 and c-MYC. The PB- and BM-derived iPSCs were indistinguishable from human embryonic stem cells (hESCs) with respect to morphology, expression of surface antigens and pluripotency-associated transcription factors, global gene expression profiles and differentiation potential in vitro and in vivo. Infected PB and BM MNCs gave rise to iPSCs in the presence of several cytokines, although transduction efficiencies were not high. We found that 5×105 PB MNCs, which corresponds to less than 1 ml of PB, was enough for the generation of several iPSC colonies. Generation of iPSCs from MNCs of non-mobilized PB, with its relative efficiency and ease of harvesting, could enable the therapeutic use of patient-specific pluripotent stem cells.

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miRNAs affected in peripheral blood CD34+ cells by stroke

Project description:To understand the differentially expressed miRNAs in the peripheral blood CD34+ cells after stroke (first 24 hours after symptom onset), we have employed mRNA sequencing. Among 45 patients included in the study, 6 patients were chosen depending on the clinical outcome (3 patients with "good" and 3 patients with "bad" clinical outcome after 3 months). Immediately after the collection of the peripheral blood from the patients, mononuclear cells (MNCs) were isolated by density centrifugation. Then, CD34+ cells were isolated from MNCs using MACS columns. The cells were immediately frozen in RNAProtect Cell Reagent. The cell samples were shipped to Seqmatic LLC (Fremont, CA, USA) for RNA isolation and miRNA sequencing. RNA integrity measurements, library preparation and next generation sequencing were conducted at Seqmatic LLC, CA, USA.

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