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New-born rat and kitten cerebellum may be maintained for prolonged periods (over 5 months) in the Maximow assembly if explanted on to a coverslip previously coated with a thin gel of reconstituted rat tail collagen and fed a glucose-enriched "natural" medium. After a 2 week period of adjustment and early outgrowth, most cultures exhibit myelin formation. Axons located within the surrounding neuroglial sheet of the explant area myelinate. The sheaths are first evident as long, unsegmented, smooth, parallel, refractile lines. Simultaneously, neuronal nuclei tend to assume central positions and powdery granules of Nissl substance and lipoid materials begin to accumulate within the cytoplasm. During prolonged maintenance, axons may increase in width and the myelin may thicken. Some exhibit transient irregularities and swellings. Degeneration of some axons occurs manifested either by (a) progressive swellings and distortions of the myelin sheath and thinning of intervening portions of the axons which finally yield, leaving the swellings as myelin bodies, or by (b) small aneurysm-like distortions of myelin sheaths on thinning axons which become dull, irregular, and thread-like filaments beaded by the former herniations. The observations are compared with previous studies of in vitro and in vivo myelin formation with particular reference to neuronal-neuroglial relationships.

Serial Observations on Patterns of Growth, Myelin Formation, Maintenance and Degeneration in Cultures of New-Born Rat and Kitten Cerebellum