Enrichment for submitotic cell populations using flow cytometry.
BACKGROUND
One of the most dramatic events during the course of the mammalian cell cycle is mitosis, when chromosomes condense and segregate, the nuclear envelope breaks down, and the cell divides into two daughter cells. Although cells undergoing mitosis are cytologically distinguishable from nonmitotic cells, few molecular markers are available to specifically identify mitotic cells, especially cells within different stages of mitosis.


METHODS
We applied the flow cytometric method of Juan et al. (Cytometry 32:71-77, 1998) to obtain cells with various levels of the molecular markers cyclin B1 and phosphorylated histone H3; fluorescence microscopy was then used to identify sorted cells in different stages of mitosis.


RESULTS
We observed the substantial enrichment of submitotic cell populations.


CONCLUSIONS
This method represents an effective approach to obtain an enriched population of submitotic cells without the use of drug treatments or prior synchronization.
Widrow
R J
RJ
Molecular and Cellular Biology Program of the University of Washington and the Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Laird
C D
CD
D015415:Biomarkers / Q000032:analysis; D002460:Cell Line; D019926:Cyclin B / Q000032:analysis; D056744:Cyclin B1; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006657:Histones / Q000032:analysis; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence; D008938:Mitosis / Q000235:genetics*; D010766:Phosphorylation
D015415:Biomarkers; C534699:CCNB1 protein, human; D019926:Cyclin B; D056744:Cyclin B1; D006657:Histones