Using intracellular markers to identify a novel set of surface markers for live cell purification from a heterogeneous hIPSC culture

Sci Rep. 2018 Jan 16;8(1):804. doi: 10.1038/s41598-018-19291-4.

Abstract

Human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) can provide sources for midbrain dopaminergic (mDA) neural progenitors (NPCs) for cell therapy to treat Parkinson's disease (PD) patients. However, the well-known line-to-cell line variability in the differentiation capacity of individual cell lines needs to be improved for the success of this therapy. To address this issue, we sought to identify mDA NPC specific cell surface markers for fluorescence activated cell sorting (FACS). Through RNA isolation after sorting for NPCs based on staining for cell-specific transcription factors followed by microarray, we identified two positive cell surface markers (CORIN and CD166) and one negative cell surface marker (CXCR4) for mDA NPC sorting. These three markers can enrich floor plate NPCs to 90% purity, and the sorted NPCs more efficiently differentiate to mature dopaminergic neurons compared to unsorted or CORIN+ alone mDA NPCs. This surface marker identification strategy can be used broadly to facilitate isolation of cell subtypes of interest from heterogeneous cultures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / analysis
  • Biomarkers / analysis*
  • Cell Adhesion Molecules, Neuronal / analysis
  • Fetal Proteins / analysis
  • Flow Cytometry / methods*
  • Human Embryonic Stem Cells / chemistry*
  • Human Embryonic Stem Cells / classification
  • Human Embryonic Stem Cells / physiology*
  • Humans
  • Induced Pluripotent Stem Cells / chemistry*
  • Induced Pluripotent Stem Cells / classification
  • Induced Pluripotent Stem Cells / physiology*
  • Membrane Proteins / analysis*
  • Receptors, CXCR4 / analysis
  • Serine Endopeptidases / analysis

Substances

  • ALCAM protein, human
  • Antigens, CD
  • Biomarkers
  • CXCR4 protein, human
  • Cell Adhesion Molecules, Neuronal
  • Fetal Proteins
  • Membrane Proteins
  • Receptors, CXCR4
  • CORIN protein, human
  • Serine Endopeptidases