Time-course Transcriptional Profiling of Human Amniotic Fluid-derived Stem Cells Using Microarray

Abstract

To maintain the homeostasis of stem cells and prevent their ability to initiate tumorigenesis, it is important to identify and modify factors that prevent or accelerate stem cell senescence. We used microarrays to attempt to identify such factors in human amniotic fluid (HAF)-derived stem cells. To identify gene expression changes over a time course, we compared gene expression profiles of HAF-derived stem cells in different passages (1(st), 2(nd), 4(th), 6(th), 8(th), and 10(th)) using a Sentrix Human illumina microarray. Of the 25,804 genes in the microarray chip, 1,970 showed an over 2-fold change relative to the control (the 1(st) passage)-either upregulated or downregulated. Quantitative real-time PCR validated the microarray data for selected genes: markedly increased genes were CXCL12, cadherin 6 (CDH6), and folate receptor 3 (FOLR3). Downregulated genes included cyclin D2, keratin 8, insulin-like growth factor 2 (IGF2), natriuretic peptide precursor B (NPPB) and cellular retinoic acid binding protein 2 (CRABP2). The expression pattern of the selected genes was consistent with the microarray data except for CXCL12 and IGF2. Interestingly, the expression of NPPB was dramatically downregulated along the time course; it was almost completely shut-down by the 10(th) passage. In contrast, FOLR3 mRNA expression was dramatically increased. Taken together, although a function for NPPB and FOLR3 in stem cell senescence has not been reported, our results strongly suggest that NPPB and/or FOLR3 play a significant role in the regulation of stem cell senescence.