Journal
BRAIN RESEARCH PROTOCOLS
Volume 16, Issue 1-3, Pages 50-57Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.brainresprot.2005.10.004
Keywords
adult DRG; cell selection; magnetic bead; surface receptor
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Current protocols for preparing primary sensory neuron cultures are inadequate when studying individual subpopulations of dorsal root ganglion (DRG) neurons. The DRG is made up of a heterogeneous population of cells, making it difficult to study treatment effects on any given population in mass cultures. Thus, we describe a procedure using magnetic beads from Dynal (TM) to select and plate viable populations of neurons based on expression of specific cell surface markers. We show that, by the use of the lectin 1134, we can select a highly enriched viable subpopulation of GDNF-responsive DRG neurons, leaving a viable population of non-selected IB4-ve, Trk+ve neurons. Key factors for successful cultures are (i) quick and careful dissection of DRGs from 4- to 5-week-old Sprague-Dawley rats, (ii) adequate removal of debris and non-neuronal contamination and (iii) gentle handling of bead-bound cells during selection. (c) 2005 Elsevier B.V. All rights reserved.
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