Reduced variability of neural progenitor cells and improved purity of neuronal cultures using magnetic activated cell sorting

PLoS One. 2019 Mar 27;14(3):e0213374. doi: 10.1371/journal.pone.0213374. eCollection 2019.

Abstract

Genetic and epigenetic variability between iPSC-derived neural progenitor cells (NPCs) combined with differences in investigator technique and selection protocols contributes to variability between NPC lines, which subsequently impacts the quality of differentiated neuronal cultures. We therefore sought to develop an efficient method to reduce this variability in order to improve the purity of NPC and neuronal cultures. Here, we describe a magnetic activated cell sorting (MACS) method for enriching NPC cultures for CD271-/CD133+ cells at both early (<2-3) and late (>10) passage. MACS results in a similar sorting efficiency to fluorescence activated cell sorting (FACS), while achieving an increased yield of live cells and reduced cellular stress. Furthermore, neurons derived from MACS NPCs showed greater homogeneity between cell lines compared to those derived from unsorted NPCs. We conclude that MACS is a cheap technique for incorporation into standard NPC differentiation and maintenance protocols in order to improve culture homogeneity and consistency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AC133 Antigen / metabolism
  • Cell Differentiation
  • Cell Line
  • Cell Separation / methods*
  • Cellular Reprogramming Techniques
  • Flow Cytometry / methods
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Magnetics
  • Nerve Tissue Proteins / metabolism
  • Nestin / metabolism
  • Neural Stem Cells / cytology*
  • Neural Stem Cells / metabolism
  • Neurons / cytology*
  • Neurons / metabolism
  • Receptors, Nerve Growth Factor / metabolism
  • SOXB1 Transcription Factors / metabolism

Substances

  • AC133 Antigen
  • NES protein, human
  • NGFR protein, human
  • Nerve Tissue Proteins
  • Nestin
  • PROM1 protein, human
  • Receptors, Nerve Growth Factor
  • SOX2 protein, human
  • SOXB1 Transcription Factors