Reconstruction of Dissociated Chick Brain Cells in Rotation-Mediated Culture

Abstract

1. By a technique of rotation-mediated cultivation, cells dissociated from brain 6- to 9-day chick embryos were cultured in vitro. Both cerebral and midbrain cells dissociated from successively older embryos made progressively smaller aggregates. It is suggested from the observation of histogenesis in the aggregates that the decrease in the size of aggregates may be related to the cellular differentiation.
2. In Eagle's basal medium containing bovine serum in various concentrations, a largest aggregates in size were formed with 10% or 20% serum. Aggregates cultured in Eagle's basal medium alone showed an approximately similar size to those cultured with 5% or 40% serum, but the former showed a sign of necrosis. These results indicate that serum seems to have no direct effect on the cell-to-cell contact, although it may be necessary for general cell metabolism.
3. Aggregates of the cerebrum, the midbrain, and the neural-retina cells from 7 1/2-day chick embryos were cultured for varying days. Aggregates of cerebral cells and midbrain cells showed a rise in size. This may be due to cell growth, or the formation of fibres. Aggregates of neural-retina cells showed changes in size produced by the fusion or fragmentation of the aggregates.
4. Commingled aggregates of cerebral and neural-retina cells showed a histogenesis in which the neural-retina tissue is arranged concentrically around the cerebral tissue. This may indicate that a sorting out took place between the cerebral cells and the neural-retina cells. As a result of these facts, the meanings of changes in aggregation patterns were discussed.
This work was aided in part by Grant No. RF-00058 (to Y. Kuroda) from the National Institutes of Health, U. S. P. H. S.. The author wishes to express his deepest appreciation to Assistant Professor Y. Kuroda, for his constant interests and advices throughout the course of this investigation and the reading of this manuscript.