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Merck

Purification of dorsal root ganglion neurons from rat by immunopanning.

Cold Spring Harbor protocols (2014-08-03)
J Bradley Zuchero
ABSTRACT

Dorsal root ganglion neurons (DRGs) are sensory neurons that facilitate somatosensation and have been used to study neurite outgrowth, regeneration, and degeneration and PNS and CNS myelination. Studies of DRGs have relied on cell isolation strategies that generally involve extended culture in the presence of antimitotic agents or other cytotoxic treatments that target dividing cells. The surviving cells typically are dependent on serum for growth. Other methods, involving purification of DRGs based on their large size, produce low yield. In contrast, the immunopanning-based method described here for prospective isolation of DRGs from rodents allows for rapid purification in the absence of antimitotic agents and serum. These DRG cultures take place in a defined medium. They are free of Schwann cells and other glia and thus can be used to study the role of glia in the biology of DRG neurons.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Cysteine hydrochloride monohydrate, reagent grade, ≥98% (TLC)
Sigma-Aldrich
Sodium selenite, γ-irradiated, lyophilized powder, BioXtra, suitable for cell culture
Sigma-Aldrich
N-Acetyl-L-cysteine, BioXtra, ≥99% (TLC)
Sigma-Aldrich
Forskolin, from Coleus forskohlii, ≥98% (HPLC), powder
Sigma-Aldrich
Earle′s Balanced Salts, With sodium bicarbonate, without calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture