TaqMan® OpenArray® high-throughput transcriptional analysis of human embryonic and induced pluripotent stem cells

Methods Mol Biol. 2013:997:191-201. doi: 10.1007/978-1-62703-348-0_15.

Abstract

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.

MeSH terms

  • Cell Culture Techniques
  • Cells, Cultured
  • DNA Probes / genetics*
  • Embryonic Stem Cells / metabolism*
  • Fibroblasts / cytology
  • Foreskin / cytology
  • Gene Expression
  • Gene Expression Profiling*
  • Humans
  • Induced Pluripotent Stem Cells / metabolism*
  • Male
  • Oligonucleotide Array Sequence Analysis*
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA Probes
  • RNA, Messenger