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Gene expression profile at a single cell level of differentiating J1 mouse embryonic stem cells cocultured with OP9 cells

ABSTRACT: The OP9 coculture system is widely used to differentiate pluripotent stem cells into mesodermal lineages. We used single cell RNA sequencing to analyze differentiation trajectories and heterogeniety of differentiating pluripotent stem cells using the OP9 coculture system.

ORGANISM(S): Mus musculus

PROVIDER: GSE271441 | GEO | 2024/07/10

REPOSITORIES: GEO

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RNA-seq analysis of mouse pluripotent stem cells derived cells

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Project description:In the bone marrow B cell development occurs in initimate and essential association with stromal cells. Known effects of stromal cells on B cell development have been shown to be mediated through direct contact and by soluble factors produced by stromal cells. Our findings suggest that cell-nonautonomous Hh signaling may play an important role in B cell lymphopoiesis. We therefore interrogated OP9 stromal cells for Hh-dependent transcripts that may confer B lymphopoietic identity. We generated OP9 stromal cells deficient in Hh signaling through targeting the non-redundant, pathway-obligate GPCR Smoothened. OP9 cells were transduced with RNAi pLKO.1-Puro lentiviral particles expressing a scrambled control shRNA (NT) or shRNA targeting Smoothened (Smo-kd). RNA from stable NT and Smo-kd OP9 cells were isolated in triplicate and global changes in transcripts were analyzed using the Affymetrix Mouse Exon 1.0 ST gene chip.

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Project description:Comparison between ex vivo immature, mature and stimulated T cells and in vitro generated counterparts. The T cells generated in vitro were cultured on OP9-DL1 stroma supplied with growth factors.

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Expression data from Hh signaling-deficient OP9 stromal cells

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