Generation of Integration-Free Patient Specific iPS Cells Using Episomal Plasmids Under Feeder Free Conditions

Methods Mol Biol. 2016:1353:355-66. doi: 10.1007/7651_2015_204.

Abstract

Reprogramming somatic cells into a pluripotent state involves the overexpression of transcription factors leading to a series of changes that end in the formation of induced pluripotent stem cells (iPSCs). These iPSCs have a wide range of potential uses from drug testing and in vitro disease modelling to personalized cell therapies for patients. While viral methods for reprogramming factor delivery have been traditionally preferred due to their high efficiency, it is now possible to generate iPSCs using nonviral methods at similar efficiencies. We developed a robust reprogramming strategy that combines episomal plasmids and the use of commercially available animal free reagents that can be easily adapted for the GMP manufacture of clinical grade cells.

Keywords: Episomal plasmids; Induced pluripotent stem cells; Reprogramming.

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Amides / pharmacology
  • Animals
  • Antigens, Surface / genetics
  • Antigens, Surface / metabolism
  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects
  • Cellular Reprogramming*
  • Dermis / cytology
  • Dermis / metabolism
  • Fibroblasts / cytology*
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Gene Expression
  • Hermanski-Pudlak Syndrome / genetics
  • Hermanski-Pudlak Syndrome / metabolism
  • Hermanski-Pudlak Syndrome / pathology*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Lewis X Antigen / genetics
  • Lewis X Antigen / metabolism
  • Mice
  • Models, Biological
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Pyridines / pharmacology
  • Serial Passage
  • Teratoma / genetics
  • Teratoma / metabolism
  • Teratoma / pathology
  • Transgenes

Substances

  • Amides
  • Antigens, Surface
  • Intercellular Signaling Peptides and Proteins
  • Lewis X Antigen
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Pyridines
  • TRA-1-81 antigen, human
  • Y 27632
  • Alkaline Phosphatase