Modeling amyotrophic lateral sclerosis in pure human iPSc-derived motor neurons isolated by a novel FACS double selection technique

Neurobiol Dis. 2015 Oct:82:269-280. doi: 10.1016/j.nbd.2015.06.011. Epub 2015 Jun 21.


Amyotrophic lateral sclerosis (ALS) is a severe and incurable neurodegenerative disease. Human motor neurons generated from induced pluripotent stem cells (iPSc) offer new perspectives for disease modeling and drug testing in ALS. In standard iPSc-derived cultures, however, the two major phenotypic alterations of ALS--degeneration of motor neuron cell bodies and axons--are often obscured by cell body clustering, extensive axon criss-crossing and presence of unwanted cell types. Here, we succeeded in isolating 100% pure and standardized human motor neurons by a novel FACS double selection based on a p75(NTR) surface epitope and an HB9::RFP lentivirus reporter. The p75(NTR)/HB9::RFP motor neurons survive and grow well without forming clusters or entangled axons, are electrically excitable, contain ALS-relevant motor neuron subtypes and form functional connections with co-cultured myotubes. Importantly, they undergo rapid and massive cell death and axon degeneration in response to mutant SOD1 astrocytes. These data demonstrate the potential of FACS-isolated human iPSc-derived motor neurons for improved disease modeling and drug testing in ALS and related motor neuron diseases.

Keywords: Amyotrophic lateral sclerosis (ALS); Axon degeneration; Cell death; Fluorescent-activated cell sorting (FACS); HB9; Induced pluripotent stem cells (iPSc); Motor neuron disease; Superoxide dismutase 1 (SOD1); p75NTR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Amyotrophic Lateral Sclerosis* / pathology
  • Amyotrophic Lateral Sclerosis* / physiopathology
  • Astrocytes / pathology
  • Astrocytes / physiology
  • Axons / pathology
  • Axons / physiology
  • Cell Survival
  • Cells, Cultured
  • Child
  • Coculture Techniques
  • Flow Cytometry / methods*
  • Genes, Reporter
  • Humans
  • Induced Pluripotent Stem Cells* / physiology
  • Lentivirus
  • Motor Neurons* / pathology
  • Motor Neurons* / physiology
  • Mutation
  • Nerve Degeneration / pathology
  • Nerve Degeneration / physiopathology
  • Nerve Tissue Proteins / metabolism
  • Receptors, Nerve Growth Factor / metabolism
  • Superoxide Dismutase / genetics
  • Superoxide Dismutase / metabolism
  • Superoxide Dismutase-1


  • NGFR protein, human
  • Nerve Tissue Proteins
  • Receptors, Nerve Growth Factor
  • SOD1 protein, human
  • Superoxide Dismutase
  • Superoxide Dismutase-1