Isolation and Culture of Rat DRG Neurons

The lumbar (L3-L5) DRG SNs were isolated as described ^{15 (link)}. Primary rat DRG neuronal cells (Lonza, R-DRG-505, Alpharetta, GA) were cultured in primary neuron growth medium (Lonza, #CC-4461) with 2% FBS, L-glutamine, gentamycin sulfate-amphotericin (GA-1000, Lonza, #CC-4083) and neural survival factor-1 (NSF-1, Lonza, #CC-4323) as instructed. Immortalized rat DRG neuronal cells, 50B11, were a generous gift from Dr. Hoke, Johns Hopkins University and cultured in Neurobasal medium (Gibco) supplemented with 10% FBS, 1 x B27 supplement (Gibco), 0.2% glucose and 0.5 mM glutamine (neuron growth medium) ^{30 (link)}. The rat DRG/mouse neuroblastoma hybrid cells, F11, were cultured as described ^{53 (link)}.

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OKUI T., Hiasa M., Hata K., Roodman G.D., Nakanishi M, & Yoneda T. (2023). The acid-sensing nociceptor TRPV1 controls breast cancer progression in bone via regulating HGF secretion from sensory neurons. Research Square.

Publication Preprint 2023

R-DRG-505 GA-1000 NSF-1 Neurobasal medium B27 supplement

Amphotericin Cells Factor 1 Gentamycin sulfate Glucose Glutamine Growth medium Hybrid cells Lumbar Mouse Neural Neuroblastoma Neuron Neuron growth Supplement

Corresponding Organization :

Other organizations : Shimane University, Tokushima University, Osaka University, Indiana University, Wakayama Medical University

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Variable analysis

independent variables

Isolation of lumbar (L3-L5) DRG sensory neurons (SNs)

dependent variables

Culturing of primary rat DRG neuronal cells
Culturing of immortalized rat DRG neuronal cells (50B11)
Culturing of rat DRG/mouse neuroblastoma hybrid cells (F11)

control variables

Culture media compositions for each cell type
Supplementation with FBS, L-glutamine, gentamycin sulfate-amphotericin (GA-1000), and neural survival factor-1 (NSF-1)

positive controls

None specified

negative controls

None specified

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