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Time series messenger RNA analysis of immature T-cells and reMAIT cells by microarrays

ABSTRACT: To explore the molecular mechanisms underlying selective differentiation of MAIT-derived iPSCs into reMAITs, we conducted mRNAs profiling based on microarrays. We sampled reMAITs during the time course of differentiation from iPSCs as well as mature MAITs isolated from cord blood (CB MAITs). As control, we used immature T cells differentiated from hematopoietic stem cells (HSCs). For each of reMAITs and the immature T cells, we selected four time points: Start, Early, Middle, and Late.

ORGANISM(S): Homo sapiens

PROVIDER: GSE87739 | GEO | 2017/03/14

SECONDARY ACCESSION(S): PRJNA347297

REPOSITORIES: GEO

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Time series micro RNA analysis of immature T-cells and reMAIT cells by microarrays

Project description:To explore the molecular mechanisms underlying selective differentiation of MAIT-derived iPSCs into reMAITs, we conducted micro RNAs profiling based on microarrays. We sampled reMAITs during the time course of differentiation from iPSCs as well as mature MAITs isolated from cord blood (CB MAITs). As control, we used immature T cells differentiated from hematopoietic stem cells (HSCs). For each of reMAITs and the immature T cells, we selected four time points: Start, Early, Middle, and Late. MAITs isolated from CB were reprogrammed to iPSCs (PMID:23523177). The MAIT-derived iPSCs were cultured on OP9, and CD34+ CD43+ cells were isolated. These CD34+ CD43+ precursor cells were furthered cultured on OP9/DL1 for re-differentiation into MAITs. Based on the observation of the reported surface antigen profiles (PMID:23523177), reMAITs were harvested at four different time points: day 0 (Start), day 4 (Early), day 7-10 (Middle), and after day 30 (Late). For the immature T cells, CD34+ cells were isolated from CB using CD34 MicroBead Kit (Miltenyi Biotech). These CD34+ HSCs were cultured on OP9/DL1 for differentiation into T cell lineage as previously described (PMID:15494433). Based on the surface antigen profiles, the immature T cells were harvested at four different time points: CD34+ cells at day 0 (Start), CD4- CD8- double negative cells at day 21 (Early) and day 40 (Midlle), and CD4+ CD8+ double positive cells after day 50 (Late). Total RNA was extracted from each sample using RNeasy Kit (Quiagen). For micro RNA analysis, RNA was labeled with Cy3-pCp by ligation, and subjected to analysis using Human miRNA Microarray Release 19.0 8x60K (Agilent).

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