Unit

UNIT 1H.7 Slice Culture Method for Studying Migration of Neuronal Progenitor Cells Derived from Human Embryonic Stem Cells (hESC)

  1. Radmila Filipovic1,
  2. Saranya Santosh Kumar1,
  3. Ben A. Bahr2,
  4. Joseph Loturco1

Published Online: 16 MAY 2014

DOI: 10.1002/9780470151808.sc01h07s29

Current Protocols in Stem Cell Biology

Current Protocols in Stem Cell Biology

How to Cite

Filipovic, R., Kumar, S. S., Bahr, B. A. and Loturco, J. 2014. Slice Culture Method for Studying Migration of Neuronal Progenitor Cells Derived from Human Embryonic Stem Cells (hESC). Current Protocols in Stem Cell Biology. 29:H:1H.7:1H.7.1–1H.7.14.

Author Information

  1. 1

    Department of Physiology and Neurobiology, University of Connecticut, Storrs, Connecticut

  2. 2

    Biotechnology Research and Training Center, William C. Friday Laboratory, University of North Carolina Pembroke, Pembroke, North Carolina

Publication History

  1. Published Online: 16 MAY 2014

Abstract

In this unit we describe an overlay brain slice culture assay for studying migration of transgenic neurospheres derived from human embryonic stem cells (hESC). Neuronal progenitor cells were generated from hESC by derivation of embryoid bodies and rosettes. Rosettes were transfected using the PiggyBac transposon system with either control plasmids (GFP) or plasmid encoding a gene important for migration of neuronal progenitor cells, Doublecortin (DCX). Transfected cells were subsequently grown in low-adhesion plates to generate transgenic human neurospheres (t-hNS). Organotypic slice cultures were prepared from postnatal rat forebrain and maintained using the interface method, before transfected t-hNS were overlaid below the cortex of each hemisphere. After 1 to 5 days, forebrain slices were fixed and processed for immunofluorescence. The distance at which cells migrated from the center of neurospheres to the host forebrain tissue was measured using Image J software. This protocol provides details for using the slice culture method for studying migration and integration of human neuronal cells into the host brain tissue. Curr. Protoc. Stem Cell Biol. 29:1H.7.1-1H.7.14. © 2014 by John Wiley & Sons, Inc.

Keywords:

  • organotypic slice cultures;
  • overlay essay;
  • human embryonic stem cell;
  • neuronal progenitors;
  • piggyBac;
  • migration