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, respective ly, is described. Five different porous microcarriers were examined fo r their capabilities to support maturation of neurons, as evaluated by determinations of protein content and glucose uptake. These systems p roved suitable for perfusion in the NMR tube for long periods of time. The energetic status of the cells was monitored by P-31-NMR, and a ve ry good temporal resolution (less than 30 min) was achieved with super fused astrocytes; thus, this provides a valuable method for real time NMR investigation of biochemical processes in primary astrocytes, Sign ificantly longer acquisition times (approximately 8 h) were required i n 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.