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The Journal of Neuroscience, March 10, 2004, 24(10):2357-2365; doi:10.1523/JNEUROSCI.4083-03.2004

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Cellular/Molecular
Efficient Isolation and Gene Expression Profiling of Small Numbers of Neural Crest Stem Cells and Developing Schwann Cells

Johanna Buchstaller,¹ Lukas Sommer,¹ Matthias Bodmer,¹ Reinhard Hoffmann,² Ueli Suter,¹ and Ned Mantei¹

¹Institute of Cell Biology, Department of Biology, Swiss Federal Institute of Technology, CH-8093 Zürich, Switzerland, and ²Max-von-Pettenkofer-Institut, D-80336 Munich, Germany

Schwann cells develop from multipotent neural crest stem cells and are important for neuronal survival, maintenance of axonal integrity, and myelination. We used transgenic mice expressing green fluorescent protein in a tissue-specific manner to isolate viable, pure populations of neural crest stem cells and developing Schwann cells, which are not readily accessible by microdissection. Starting with the minute amounts of RNA obtained, a two-round amplification procedure was used to achieve reproducible DNA array hybridizations. We validated our screening procedure by comparisons with the literature and by in situ hybridization. Stage-to-stage comparisons and hierarchical clustering for neural crest and five stages of Schwann cell development suggest a wealth of candidates for genes involved in stem cell regulation and in early Schwann cell development. The combination of methods applied in this study should be generally useful for isolating and profiling other stem cell and difficult to isolate cell populations.

Key words: development; gene; glia; neural tube neural crest; neural plate; Schwann cell; sensory neurons; green fluorescent protein; GFP; fluorescence-activated cell sorting; FACS; peripheral nervous system; PNS; microarray; expression analysis

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Received Sep 4, 2003; revised December 30, 2003; accepted January 8, 2004.

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