Isolation and long-term survival of adult human sensory neurons in vitro.
OBJECTIVE
To determine whether adult human dorsal root ganglion neurons can be isolated and maintained in long-term tissue culture, where they would extend processes.


METHODS
Dorsal root ganglia were removed from adult human organ donors within 2 hours of clamping the aorta. They were then treated with enzymes for one hour, triturated to dissociate the neurons and their satellite cells, and the individual neurons were then plated in tissue culture dishes in medium containing serum.


RESULTS
Isolated adult human dorsal root ganglion neurons survive in vitro for more than 2 1/2 months, in the absence of exogenously supplied neurotrophins. where they remain electrically excitable and extend processes,


CONCLUSIONS
Isolated adult human dorsal root ganglion neurons survive in culture for more than 2 1/2 months, extend processes, and remain electrically excitable, without exogenous neurotrophins. These results suggest that, adult human sensory neurons do not require exogenous neurotrophins for survival and process outgrowth, or that sufficient factors were provided by the small number of satellite cells in the cultures. In addition, the neurons survive well in spite of an initial period of up to 14 hours of hypoxia, between the time the aorta was clamped and when the plated neurons were placed in an incubator with the appropriate O2/CO2 environment.
Sosa
I J
IJ
Institute of Neurobiology, Medical Sciences Campus, University of Puerto Rico, San Juan.
Reyes
O
O
Inserni
J
J
Kuffler
D P
DP
D000200:Action Potentials / Q000502:physiology; D000328:Adult; D002448:Cell Adhesion / Q000502:physiology; D002469:Cell Separation; D002470:Cell Survival; D046508:Culture Techniques; D004558:Electric Stimulation; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D008839:Microelectrodes; D008875:Middle Aged; D009475:Neurons, Afferent / Q000166:cytology / Q000502:physiology*; D013997:Time Factors