Enrichment of spinal cord cell cultures with motoneurons.

摘要: Spinal cord cell cultures contain several types of neurons. Two methods are described for enriching such with motoneurons (defined here simply as cholinergic cells that capable innervating muscle). In the first method, 7-day embryonic chick spinal neurons were separated according to size by 1 g velocity sedimentation. It is assumed among largest present at this stage. The cords dissociated vigorously so 95-98% in initial suspension isolated from one another. Cells leading fractions (large fractions: LCFs) about seven times much choline acetyltransferase (CAT) activity per unit cytoplasm do trailing (small SCFs). Muscle seeded LCFs develop 10-70 CAT SCFs and six control (unfractionated) cells. More than 20% large LCF-muscle innervate nearby myotubes. second gently 4-day maintained vitro. This approach based on earlier observations withdraw form mitotic cycle developing embryo (Hamburger, V. 1948. J. Comp. Neurol. 88:221-283; Levi-Montalcini, R. 1950. Morphol. 86:253-283). 4-Day cord-muscle three plates, prepared same (gentle) manner. 50% relatively Thus, both useful steps toward complete isolation motoneurons. Both should facilitate study development nerve-muscle synapse formation.