A simple reproducible method for prometaphase chromosome analysis.
A method is described for the analysis of chromosomes in prophase and early metaphase. It involves culturing the lymphocytes in medium RPMI-1640, supplemented with 10% autologous plasma instead of fetal bovine serum. Living cells are treated with actinomycin D and colcemid for 1 h prior to harvest and harvested early at 65 h of incubation, using a hypotonic solution formulated by Ohnuki (1968). The method has been tested on several hundred clinical samples on a routine basis. On average, 30% of the dividing cells were in prometaphase.
D001399:Azure Stains; D002455:Cell Division; D002871:Chromosome Banding; D002877:Chromosomes, Human / Q000648:ultrastructure*; D003078:Colchicine; D003470:Culture Media; D003609:Dactinomycin; D006801:Humans; D006982:Hypertonic Solutions; D008214:Lymphocytes / Q000166:cytology*; D008677:Metaphase; D010949:Plasma; D011418:Prophase
D001399:Azure Stains; D003470:Culture Media; D006982:Hypertonic Solutions; D003609:Dactinomycin; D003078:Colchicine