Human clonogenic cells in vitro. I. Improved preparatory techniques for the collection and concentration of leukemic colonies from methylcellulose cultures for morphologic, cytokinetic, and histochemical evaluation.
We have developed simple methods for the rapid isolation and concentration of human leukemic colonies for cytologic evaluation. Colonies, grown in a medium containing methylcellulose, are gently washed from their culture dishes after reducing the viscosity of the colony layer. Colonies are washed and concentrated intact by mild centrifugation (10-20 g/5 min). Once isolated, colonies may enter any number of procedures for morphologic and cytokinetic analyses including exposure to radiolabeled compounds, cytocentrifugation, reaction with antibodies specific for certain cell surface markers, and fixation for routine light and electron microscopy. In addition, suspended or smeared colonies may be exposed to various substrates for the demonstration of endogenous enzymatic markers. Examples of possible preparations are presented.
D002469:Cell Separation / Q000379:methods*; D002471:Cell Transformation, Neoplastic / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D002478:Cells, Cultured; D002999:Clone Cells / Q000166:cytology / Q000473:pathology; D006410:Hematopoiesis; D006801:Humans; D007399:Interphase; D007943:Leukemia, Hairy Cell / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D008747:Methylcellulose
D008747:Methylcellulose