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Human Pluripotent Stem Cell Naïve State qPCR Array

For characterization of gene expression associated with hPSC naïve or primed states

概要
技术资料
数据及文献

概要

The Human Pluripotent Stem Cell (hPSC) Naïve State Quantitative Polymerase Chain Reaction (qPCR) Array is designed for characterization of hPSCs and their status in the spectrum from naïve to primed pluripotency. Naïve state hPSCs are self-renewing, retain the properties of the pre-implantation blastocyst, can differentiate to all somatic lineages without lineage bias, and show potential to differentiate into the germ lineage. Primed hPSCs are also self-renewing and can contribute to all somatic lineages, however some lines show lineage-specific differentiation bias. This qPCR array provides the gene expression profile of primed and naïve state hPSCs. Genes were selected based on their demonstrated differential expression in primed and naïve state hPSCs (Chan et al.; Davidson et al.; Gafni et al.; Theunissen et al.; Takashima et al.) or in hPSC-derived early ectodermal, endodermal, and mesodermal lineage cells (Huang et al.).

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 naïve and primed PSCs and their derivatives, as well as 6 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

数据及文献

Data

Figure 1. Human Pluripotent Stem Cell Naïve State qPCR Array Plate Configuration

Figure 2.

Quantitative PCR analysis of gene expression of H1 hESCs cultured in a naive-like state compared to H1 hESCs cultured under a primed state. Genes associated with higher expression in a naive state than a primed state are shown in light green, genes expected to be more highly expressed in a primed state compared to a naive state are shown in orange and pluripotency associated genes expressed highly in both naive and primed states are shown in yellow. Genes associated with endoderm (red), mesoderm (light blue) ectoderm (dark green) and partially reprogrammed genes (dark blue) in feeder-free versus feeder-dependent tissue culture are also show. Y-axis measures the log change expression difference and X-axis indicates the name of the gene. Experiments were run on a Viia7 Fast (Applied Biosystems) Instrument and data analysis was performed using STEMCELL qPCR analysis program (www.stemcell.com/qPCRanalysis).