Reaggregation of fetal rat brain cells in a stationary culture system I: Methodology and cell identification |
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Authors: | Rolf Bjerkvig Sverre K Steinsvåg Ole Didrik Laerum |
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Institution: | (1) The Gade Institute, Department of pathology, University of Bergen, N-5016 Haukeland Hospital, Bergen, Norway |
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Abstract: | Summary A stationary tissue culture system for reaggregation cultures of rat brain cells is described. Aggregates were formed by placing
cells at high concentrations in liquid overlay cultures on a nonadherent nutrient agar surface. No physical stress in the
form of rotation or shaking was applied to the aggregating cell population.
Transmission electron microscopy and immunohistochemistry showed that the cells developed from homogeneously dispersed, immature
cells in Day 4 aggregates, to mature astrocytes, oligodendrocytes, and neurons in Day 20 aggregates. Twenty days and older
aggregates had a tightly packed neuropil which was most prominent in a cell-sparse outer layer of the aggregates. When the
aggregates were allowed to adhere to a substrate, both glial fibrillary acidic protein (GFAP) positive and negative cells
were observed migrating out from the aggregates. Cells giving a positive reaction for neuron specific enolase (NSE) were also
present. This reaggregation procedure, with transfer of selected brain cell aggregates into agar-coated multiwells is an alternative
three-dimensional culture system which can be potentially useful in the study of morphogenesis and cell interactions in the
nervous system.
This project was supported by the Norwegian Cancer Society. |
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Keywords: | reaggregation fetal rat brain cells stationary tissue culture cell differentiation |
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