Efficient Production of Photoreceptor Precursor Cells from Human Embryonic Stem Cells

Methods Mol Biol. 2016:1307:357-69. doi: 10.1007/7651_2013_57.

Abstract

Transplantation of photoreceptor precursor cells (PPCs) differentiated from human embryonic stem cells (hESCs) is a promising approach to treat common blinding diseases such as age-related macular degeneration and retinitis pigmentosa. However, existing PPC generation methods are inefficient. To enhance differentiation protocols for rapid and high-yield production of PPCs, we focused on optimizing the handling of the cells by including feeder-independent growth of hESCs, using size-controlled embryoid bodies (EBs), and addition of triiodothyronine (T3) and taurine to the differentiation medium, with subsequent removal of undifferentiated cells via negative cell-selection. Our novel protocol produces higher yields of PPCs than previously reported while reducing the time required for differentiation, which will help understand retinal diseases and facilitate large-scale preclinical trials.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects
  • Collagen / pharmacology
  • Drug Combinations
  • Flow Cytometry
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / drug effects
  • Humans
  • Laminin / pharmacology
  • Magnetic Phenomena
  • Photoreceptor Cells / cytology*
  • Photoreceptor Cells / drug effects
  • Proteoglycans / pharmacology
  • Real-Time Polymerase Chain Reaction

Substances

  • Drug Combinations
  • Laminin
  • Proteoglycans
  • matrigel
  • Collagen