Immobilization of primary astrocytes and neurons for online monitoring of biochemical processes by NMR.
Immobilization of primary astrocytes and cerebellar granule neurons in basement membrane gel threads and on porous microcarriers, respectively, is described. Five different porous microcarriers were examined for their capabilities to support maturation of neurons, as evaluated by determinations of protein content and glucose uptake. These systems proved suitable for perfusion in the NMR tube for long periods of time. The energetic status of the cells was monitored by 31P-NMR, and a very good temporal resolution (less than 30 min) was achieved with superfused astrocytes; thus, this provides a valuable method for real time NMR investigation of biochemical processes in primary astrocytes. Significantly longer acquisition times (approximately 8 h) were required in order to obtain a spectrum of neurons with a reasonably good signal to noise ratio, even so, the results presented here and obtained with this difficult system are very encouraging.
Alves
P M
PM
Universidade Nova de Lisboa, Instituto de Tecnologia Quìmica e Biológica, Oeiras, Portugal.
Flögel
U
U
Brand
A
A
Leibfritz
D
D
Carrondo
M J
MJ
Santos
H
H
Sonnewald
U
U
D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001485:Basement Membrane; D018914:Cells, Immobilized; D003584:Cytological Techniques; D001330:Electronic Data Processing; D005240:Feasibility Studies; D005782:Gels; D009682:Magnetic Resonance Spectroscopy; D008863:Microspheres; D009474:Neurons / Q000378:metabolism*; D051381:Rats
D005782:Gels