Conversion of monkey fibroblasts to transplantable telencephalic neuroepithelial stem cells

Biomaterials. 2016 Jan:77:53-65. doi: 10.1016/j.biomaterials.2015.10.079. Epub 2015 Oct 31.

Abstract

Non-human primates provide optimal models for the development of stem cell therapies. Although somatic cells have been converted into neural stem/progenitor cells, it is unclear whether telencephalic neuroepithelial stem cells (NESCs) with stable properties can be generated from fibroblasts in primate. Here we report that a combination of transcription factors (Oct4, Sox2, Klf4) with a new culture medium induces rhesus monkey fibroblasts into NESCs, which can develop into miniature neural tube (NT)-like structures at a cell level. Furthermore, single induced NESCs (iNESCs) can generate later-stage 3D-NTs after grown on matrigel in suspension culture. iNESCs express NT cell markers, have a unique gene expression pattern biasing towards telencephalic patterning, and give rise to cortical neurons. Via transplantation, single iNESCs can extensively survive, regenerate myelinated neuron axons and synapse structures in adult monkey striatum and cortex, and differentiate into cortical neurons. Successful transplantation is closely associated with graft regions and grafted cell identities. The ability to generate defined and transplantable iNESCs from primate fibroblasts under a defined condition with predictable fate choices will facilitate disease modeling and cell therapy.

Keywords: Cortical neuron; Monkey fibroblasts; Neural regeneration in vivo; Neural tubes; Telencephalic neuroepithelial stem cells; Transdifferentiation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Lineage / drug effects
  • Cell Transdifferentiation
  • Cells, Cultured
  • Cerebral Cortex / cytology
  • Corpus Striatum / cytology
  • Culture Media / pharmacology
  • Fibroblasts / cytology*
  • Fibroblasts / drug effects
  • Genes, Reporter
  • Genetic Vectors / genetics
  • Graft Survival
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors / genetics
  • Kruppel-Like Transcription Factors / physiology
  • Lentivirus / genetics
  • Macaca mulatta
  • Male
  • Nerve Fibers, Myelinated / ultrastructure
  • Nerve Tissue Proteins / biosynthesis
  • Nerve Tissue Proteins / genetics
  • Neural Stem Cells / cytology*
  • Neural Stem Cells / transplantation
  • Neural Tube / cytology
  • Neuroepithelial Cells / cytology*
  • Neuroepithelial Cells / transplantation
  • Neurogenesis / drug effects*
  • Neurons / cytology
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / physiology
  • Organ Specificity
  • Prefrontal Cortex / cytology
  • Retroviridae / genetics
  • SOXB1 Transcription Factors / genetics
  • SOXB1 Transcription Factors / physiology
  • Telencephalon
  • Transcriptome
  • Transduction, Genetic

Substances

  • Culture Media
  • KLF4 protein, human
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors
  • Nerve Tissue Proteins
  • Octamer Transcription Factor-3
  • SOXB1 Transcription Factors