Elsevier

Brain Research

Volume 130, Issue 1, 8 July 1977, Pages 1-12
Brain Research

The maintenance and identification of mouse cerebellar granule cells in monolayer culture

https://doi.org/10.1016/0006-8993(77)90838-1Get rights and content

Summary

Methods are described for maintaining postnatal mouse cerebellar cells in monolayer culture, and for identifying granule cells in such cultures. Cells from cerebella of 7-day-old mice are dissociated with trypsin and DNAse, then plated at 1–1.5 × 106 cells/35 mm dish in a high-potassium modifications of Hams F12 medium plus 10% fetal calf serum. Under these conditions, cells grow either singly or in small clumps, and develop complex meshes of single fibers and fiber bundles over a period of several days. Granule cells are identified by a combination of several criteria including their size, shape and relative proportion of the total cell population as determined by phase contrast and scanning electron microscopy; nuclear morphology, demonstrated by transmission electron microscopy, and failure to take up [3H]γ-aminobutyric acid (GABA) in the presence of several other cells types which do, shown by autoradiography.

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