title: Effect of enzymatic and mechanical methods of dissociation on neural progenitor cells derived from induced pluripotent stem cells.
abstract: OBJECTIVE
  To determine the most effective method of dissociating neural stem and progenitor cells into a single-cell suspension.
  
  
  METHODS
  Induced pluripotent stem cells were differentiated toward the neural fate for 4 weeks before clusters were subjected to enzymatic (Accutase, trypsin, TrypLE, dispase, or DNase I) or mechanical (trituration with pipettes of varying size) or combined dissociation. Images of cells were analyzed for cluster size using ImageJ.
  
  
  RESULTS
  Cells treated with the enzymes Accutase, TrypLE, or trypsin/EDTA, these enzymes followed by trituration, or a combination one of these enzymes followed by incubation with another enzyme, including DNase I, were more likely to be dissociated into a single-cell suspension.
  
  
  CONCLUSIONS
  Cells treated with enzymes or combinations of methods were more likely to be dissociated into a single-cell suspension.
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Lindsey D
authors.initials: LD
authors.lastname: Jager
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Claire-Marie A
authors.initials: CM
authors.lastname: Canda
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Crystal A
authors.initials: CA
authors.lastname: Hall
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Cassandra L
authors.initials: CL
authors.lastname: Heilingoetter
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Joann
authors.initials: J
authors.lastname: Huynh
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Susanna S
authors.initials: SS
authors.lastname: Kwok
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Jin H
authors.initials: JH
authors.lastname: Kwon
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Jacob R
authors.initials: JR
authors.lastname: Richie
authors.affiliation: Department of Neurology, University of Wisconsin-Madison, Madison, WI, USA. Electronic address: jensen@neurology.wisc.edu.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Jensen
mesh_terms: D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D004798:Enzymes / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D058953:Neural Stem Cells / Q000166:cytology*; D010316:Particle Size; D013314:Stress, Mechanical
chemical_list: D004798:Enzymes
keywords: Dissociation; Neural progenitor cells; Single-cell suspension; Stem cells