Open AccessDifferentiation of HL‐60 cells: Cell volume and cell cycle changes
Author(s) -
Zucker Robert M.,
Whittington Karen,
Price Brandon J.
Publication year - 1983
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990030605
Subject(s) - cell cycle , flow cytometry , retinoic acid , cell , incubation , microbiology and biotechnology , cell culture , chemistry , cellular differentiation , biology , biochemistry , genetics , gene
HL‐60 promyelocytic leukemic cells can differentiate into more mature myeloid cells with the addition of dimethylsulfoxide, butyric acid or retinoic acid and can differentiate into macrophages with the addition of phorbol ester 12‐0‐tetradecanoylphorbol‐13‐acetate (TPA). After the addition of an inducer, the HL‐60 cell volume shows a daily decrease while the cell number increases at a rate similar to the untreated control cells. Flow cytometry measurements show an increase in G 1 cells and a decrease in S cells after day 1. Since the generation time is constant, the data suggest that the length of time spent in the different cell cycle stages has changed during differentiation. Within 3 hours after the addition of TPA to HL‐60 cells, selective adhesion of G 1 cells occurs. Smaller sized cells are recovered from the flask bottom and larger sized cells are recovered from the supernate. Flow cytometric analysis reveals a G 1 and S block in cells obtained from both the supernatant and from the flask bottom. After 1 day of TPA incubation, there is preferential adhesion of G 1 and G 2 cells with the nonadherent cells being primarily in the S and G 2 cell cycle stages and undergoing a cell cycle traverse.