Microglia develop in cultures initiated from disaggregated neopallial cells of newborn C,H/HeJ mice when the cultures are subjected to nutritional deprivation for 10 or more days (Ha0 et al: Int J Dev Neurosci 9:l-14,1991). In the present experiments, the cultures were pulsed with BrdU for 3 hours a
Cell cycle time of murine neopallial cells in vitro
β Scribed by J.P. Novak; S. Fedoroff
- Publisher
- John Wiley and Sons
- Year
- 1996
- Tongue
- English
- Weight
- 996 KB
- Volume
- 45
- Category
- Article
- ISSN
- 0360-4012
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β¦ Synopsis
Disaggregated glial cells from newborn CDl mouse neopallia were cultured in low concentration (4.2 x id cells/cm2) for 72 hr and then either pulse labeled with BrdU by one 2-hr pulse at various times of culturing or continuously labeled for various lengths of time. At the end of incubation, the cells were fixed and immunoreacted with BrdU. All BrdU+ and BrdUcell nuclei were counted in an area of 4.84 cm2. A three-compartment model for interpretation of the experimental data was developed consisting of active proliferating cells, non-active cells with proliferating potential, and nonproliferating cells. The model is based on assumptions of time invariance of culture conditions, random re-entry of cells into cell cycle and random exit from the proliferating pool. Furthermore, it is assumed that average values are representative for describing the numbers of cells in specific compartments as functions of time. A set of relationships representing the numbers of labeled cells for pulse labeling and continuous labeling assays is derived from these assumptions and the generally accepted representation of cell progress through the cell cycle, i.e., a genetically predetermined sequence of post-mitosis rest phase, S-phase, pre-mitosis rest phase, and mitosis. These relationships are used to evaluate the S-phase time T~ and cell cycle time T, of proliferating cells. Under our particular conditions, we obtain approximately T~ = 8 hr and T, = 16 hr, respectively. The applicability of the model and possible distorting factors are discussed.
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