Project description:Retinoic acid RA time course experiment: H1 cells(WiCell Research Institute, WA01, Passage40) maintained in TeSR1 were treated with 1uM retinoic acid (RA) in a detailed time course:day0,day0.5,day1,day1.5,day2,day3, day4 and day5 . Total RNA was prepared by usage of the Qiagen RNA extraction kit. mRNA was purified and then labeled and hybridized to HG17 NimbleGen human expression chips to evaluate changes in gene expression during RA-induced differentiation. Keywords: Retionic acid gene expression time series human embryonic stem cells
Project description:Retinoic acid RA time course experiment: H1 cells(WiCell Research Institute, WA01, Passage40) maintained in TeSR1 were treated with 1uM retinoic acid (RA) in a detailed time course:day0,day0.5,day1,day1.5,day2,day3, day4 and day5 . Total RNA was prepared by usage of the Qiagen RNA extraction kit. mRNA was purified and then labeled and hybridized to HG17 NimbleGen human expression chips to evaluate changes in gene expression during RA-induced differentiation. Keywords: Retionic acid gene expression time series human embryonic stem cells Samples from eight time points were taken after RA treatment (0,0.5,1,1.5,2,3,4,5 days) of H1 hES cells in TeSR1 medium. In addition, the experiment includes biological replicates at days 0 and 5.
Project description:There are a total of four samples each for this analysis. Each sample consists of the cells grown on three 10 cm culture plates. Each plate should have 2x106 cells for a total of 6x106 cells per sample when all three plates are combined. The first sample is undifferentiated human embryonic stem cells, the second sample is human glutamatergic neurons derived from those human embryonic stem cells, the third sample is undifferentiated human induced pluripotent stem cells and the fourth sample is human glutamatergic neurons derived from those human induced pluripotent stem cells.
Project description:The aim of this project is to differentiate human embryonic stem cells to an extra-embryonic fate, specifically the hypoblast. This is of uttermost importance given the current lack of human hypoblast stem cells.
We hypothesized that the pluripotent characteristics of the starting human embryonic stem cell population may dictate the competency for extra-embryonic cell fate specification. Based on this hypothesis and using human embryonic stem cells maintained in different naïve-like culture regimes, we have now developed conditions that allow the differentiation of human embryonic stem cells to a stable GATA6+ SOX2- population. This suggests that these cells may be putative human hypoblast stem cells. To validate this finding here we propose to perform RNA sequencing experiments of the differentiated human embryonic stem cells. By comparing their RNA expression profile to the single cell sequencing data of the human embryo that we are currently generating, we will be able to determine the identity of our GATA6+ SOX2- cells, and establish whether they represent the in vivo human hypoblast.
