Visualization of cell cycling by an improvement in slice culture methods.
Slice culture combined with the use of fluorescent dyes and/or the introduction of fluorescent protein genes provides live and three-dimensional information on cytogenetic and histogenetic events at the level of the individual cell. Using slices prepared from midembryonic mouse cerebral wall tissue upon which fine DiI crystals were placed on the pial or ventricular surface, we recently found that dividing progenitor cells do not lose their pia-connected (basal) processes and that the processes are inherited by daughter cells, including neurons (Miyata et al. [2001] Neuron 31:727-741). To understand more fully the biological significance of this inheritance process, the fate of each daughter cell should be monitored over a culture period extended long enough to allow a neuron to migrate up to the cortex or for a progenitor to proceed to the next round of division. Exposure of slices to 40%, instead of 20%, O(2) significantly improved their overall thickening, cell production, and layer formation and also provided better spatial resolution by preventing the loss of transparency that accompanies cell death.
Laboratory for Cell Culture Development, Advanced Technology Development Center, Brain Science Institute, RIKEN, Wako, Saitama, Japan. tmiyata@brain.riken.go.jp
Takaki
T
Miyata
Ayano
A
Kawaguchi
Kanako
K
Saito
Hiroshi
H
Kuramochi
Masaharu
M
Ogawa
D000818:Animals; D002232:Carbocyanines; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002540:Cerebral Cortex / Q000166:cytology* / Q000196:embryology*; D005456:Fluorescent Dyes; D051379:Mice; D008813:Mice, Inbred ICR; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology; D009924:Organ Culture Techniques / Q000379:methods*; D010100:Oxygen / Q000494:pharmacology; D010841:Pia Mater / Q000166:cytology / Q000196:embryology; D013234:Stem Cells / Q000166:cytology*
C035005:3,3'-dihexadecylindocarbocyanine; D002232:Carbocyanines; D005456:Fluorescent Dyes; D010100:Oxygen