Description
The Human Pluripotent Stem Cell (hPSC)-Derived Endoderm Quantitative Polymerase Chain Reaction (qPCR) Array is designed for characterization of hPSC-derived definitive endodermal progenitor cells and their differentiated progeny including pancreatic, hepatic, and intestinal cells. This qPCR array provides the gene expression profile of hPSC-derived endodermal cells following in vitro differentiation, and can aid in development of protocols for directed differentiation towards endodermal lineages. Genes were selected based on their demonstrated expression in hPSCs, or in hPSC-derived ectodermal, mesodermal, and endodermal lineage cells (D’Amour et al.), including pancreatic (Kroon et al.), hepatic (Hay et al.), and intestinal progenitor cells (Grapin-Botton & Melton).
qPCR is used to determine changes in steady-state mRNA levels of gene expression across multiple samples, generally normalized to the relative expression of internal control genes. Gene-specific primers amplify target sequences within cDNA pools reverse-transcribed from mRNA and, as with TaqMan® technology, hybridized sequence-specific probes provide a fluorescent signal from the 5' exonuclease activity of Taq DNA polymerase. The accumulation rate of the fluorescent signal is used to quantify the sample cDNA, and thereby the amount of mRNA in the original cell lysate.
This qPCR array contains validated primers and probes for detection of 90 genes whose expression is correlated with hPSCs or hPSC-derived ectodermal, mesodermal, and endodermal cells, or pancreatic, hepatic, and intestinal progenitor cells, as well as six endogenous (housekeeping) control genes. TBP (TATA box-binding protein) qPCR Control Template is provided as a synthetic DNA positive control. Additional gene information and a qPCR analysis tool are available at www.stemcell.com/qPCRanalysis.
