The Characteristics of Murine iPS Cells and siRNA Transfection Under Hypoxia

Methods Mol Biol. 2016:1357:465-74. doi: 10.1007/7651_2014_158.

Abstract

iPS cells are attractive for the regenerative medicine. The creation of pluripotent cells from somatic cells has great potential for basic and clinical research and application. Retroviral transduction of four or three transfection factors has been shown to initiate a reprogramming process. Here, we describe the effect of transcription factors regarding the growth and differentiation of mouse iPS cells in normoxia or hypoxia. Furthermore, we introduce the function of hypoxia-inducible factors (HIFs) in mouse iPS cells in hypoxia using RT-PCR and western blotting together with HIFs knockdown techniques.

Keywords: HIFs; Hypoxia; Murine iPS cells; Pluripotency; Transcription factors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis Regulatory Proteins
  • Basic Helix-Loop-Helix Transcription Factors / antagonists & inhibitors
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / physiology*
  • Blotting, Western / methods
  • Cell Culture Techniques / methods*
  • Cell Hypoxia
  • Cells, Cultured
  • Cellular Reprogramming Techniques / methods*
  • Cellular Reprogramming*
  • Gene Expression Regulation
  • Hypoxia-Inducible Factor 1, alpha Subunit / antagonists & inhibitors
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / physiology*
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Mice
  • Oxygen / pharmacology
  • Oxygen / physiology
  • RNA Interference*
  • RNA, Small Interfering / genetics*
  • Real-Time Polymerase Chain Reaction / methods
  • Repressor Proteins
  • Transcription Factors / antagonists & inhibitors
  • Transcription Factors / biosynthesis
  • Transcription Factors / genetics
  • Transcription Factors / physiology*
  • Transfection / methods*

Substances

  • Apoptosis Regulatory Proteins
  • Basic Helix-Loop-Helix Transcription Factors
  • Hif3a protein, mouse
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • RNA, Small Interfering
  • Repressor Proteins
  • Transcription Factors
  • endothelial PAS domain-containing protein 1
  • Oxygen