Isolation of fetal mouse motor neurons on discontinuous Percoll density gradients.
The spinal cords of fetal NIH:CR mice, gestational age Day 12 to 14, were dissected free of meninges and dorsal root ganglia, chemically dissociated, and layered onto discontinuous Percoll gradients at densities 1.040, 1.050, and 1.060 g/ml. After centrifugation (800 Xg for 15 min at 4 degrees C), three morphologically, biochemically, and immunohistologically distinct cell populations were collected from the gradient interfaces. The first interface, located at a density of 1.040 g/ml, was choline acetyltransferase enriched (0.86 +/- 0.08) compared to the second and third fractions (0.42 +/- 0.01 and 0 pmol acetylcholine synthesized/microgram protein, respectively). When simultaneously cultured with fetal mouse cardiac muscle on a gelatin-polylysine-laminin substrate in serum-free medium, these cells developed the characteristics of motor neurons.
Strong
M J
MJ
Laboratory of Central Nervous System Studies, National Institute of Neurological Disorders and Stroke, Bethesda, Maryland 20892.
Garruto
R M
RM
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D005333:Fetus / Q000294:innervation*; D007797:Laminin; D051379:Mice; D008815:Mice, Inbred Strains; D009046:Motor Neurons / Q000166:cytology*; D011205:Povidone; D012822:Silicon Dioxide
D007797:Laminin; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone