The Characteristics of Human iPS Cells and siRNA Transfection Under Hypoxia

Methods Mol Biol. 2022:2454:657-664. doi: 10.1007/7651_2020_299.

Abstract

The characteristics of pluripotent cells have great potential for basic and clinical research and application. We describe the effect of normoxia or hypoxia regarding the proliferation and pluripotency of human iPS cells using colony number counting and real-time polymerase chain reaction (PCR). In addition, the function of hypoxia-inducible factors (HIFs) in human iPS cells under hypoxic conditions is evaluated in relation to the expression of pluripotency markers by siRNA and real-time PCR. Furthermore, we introduce the change of HIF-2α expression when signal transducer and activator of transcription 3 (STAT3) is suppressed by its inhibitor, Stattic or S31 201, using RT-PCR.

Keywords: HIFs; Human iPS cells; Hypoxia; Pluripotency; STAT3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Hypoxia
  • Cell Proliferation
  • Humans
  • Hypoxia
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Induced Pluripotent Stem Cells* / metabolism
  • RNA, Small Interfering / metabolism
  • Transfection

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • RNA, Small Interfering