TaqMan® OpenArray® High-Throughput Transcriptional Analysis of Human Embryonic and Induced Pluripotent Stem Cells
It is widely accepted that somatic cells can be reprogrammed by a set of transcription factors to become embryonic stem cell-like: These reprogrammed cells, induced pluripotent stem cells (iPSCs), are nearly identical to embryonic stem cells (ESCs), because both have the capacity to self-renew and to form all cellular lineages of the body. Transcriptional differences between ESCs, iPSCs, and fibroblasts can be analyzed by quantitative PCR (qPCR) using TaqMan® Gene Expression assays, a widely used tool for rapid analysis of different cell types. In this chapter, we describe the OpenArray® platform which generates qPCR data from high-throughput instrumentation. We examined the gene signature profiles of ESCs, fibroblasts, and iPSCs with a TaqMan® OpenArray® Human Stem Cell Panel containing 631 TaqMan® Gene Expression assays that represent pathways involved in self-renewal, pluripotency, lineage patterning, transcriptional networks, stem cell differentiation, and development.
Key wordsReprogramming Fibroblast Induced pluripotent stem cells iPSC TaqMan RT-PCR Pluripotency Self-renewal Embryonic stem cells Adult stem cells
The authors thank Rene Quintanilla and Uma Lakshmipathy for providing the cells used in this study, Doug Roberts for assay selection, Vanee Pho for technical guidance and documentation, and Caifu Chen and David Keys for support and helpful discussions.
- 3.Seki T, Yuasa S, Oda M, Egashira T, Yae K, Kusumoto D, Nakata H, Tohyama S, Hashimoto H, Kodaira M, Okada Y, Seimiya H, Fusaki N, Hasegawa M, Fukuda K (2010) Generation of induced pluripotent stem cells from human terminally differentiated circulating T cells. Cell Stem Cell 7:11–14PubMedCrossRefGoogle Scholar
- 4.Zhang X, De Los Angeles A, Zhang J (2010) The art of human induced pluripotent stem cells: the past, the present and the future. Open Stem Cell J 2:2–7Google Scholar