Document Detail


Growth of single HL60 cells in liquid culture: analysis of the influences of differentiative agents.
MedLine Citation:
PMID:  8960107     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Treatment of serum-free grown HL60 cells with certain combined amounts of retinoic acid (9-cis or all-trans RA) and 1 alpha 25 dihydroxyvitamin D3 (D3) results in differentiation of 71-77% of cells towards either neutrophils or monocytes. Studies of the differentiation of HL60 cells in flask cultures does not reveal: (i) the extent to which selective growth of cells might have occurred; and (ii) the overall level of cell survival. This information can be obtained by monitoring the effects of differentiative agents on individual cells. Serum-free grown HL60 cells were cultured as single cells in microtitre wells in conditioned medium obtained from exponentially growing and serum-free cultures of HL60. This resulted in a cloning efficiency of 85% and HL60 cells doubled every 24 h. During a period of exponential growth < 0.5 to 2% of the cells generated died. Single HL60 cells were treated with 9-cis and all-trans RA (5 x 10(-7) M) together with a small amount of D3 (3.9 x 10(-14) M) to promote neutrophil differentiation. D3 alone (10(-7) M) and D3 (5 x 10(-9) M) in combination with 9-cis RA (10(-8) M) were used to promote monocyte differentiation. The growth kinetics of HL60 cell cultures that were differentiating to neutrophils and to monocytes were similar. Single-cell experiments have revealed that: (i) differentiating HL60 cells undergo a variable number of divisions (two to five) prior to arresting their growth; and (ii) up to 33% of the cells that are generated (by day 5) die. Seventy to eighty per cent of the cells in each of the wells had matured. These findings have important implications in regard to whether retinoids and D3 provide signals that determine the choice of maturation pathway or that merely facilitate selective survival and/or expansion of cells that have independently determined their differentiation fates.
Authors:
L A Wallington; J Durham; C M Bunce; G Brown
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Leukemia research     Volume:  20     ISSN:  0145-2126     ISO Abbreviation:  Leuk. Res.     Publication Date:  1996 Oct 
Date Detail:
Created Date:  1997-01-02     Completed Date:  1997-01-02     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7706787     Medline TA:  Leuk Res     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  821-9     Citation Subset:  IM    
Affiliation:
Department of Immunology, Medical School, University of Birmingham, UK.
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MeSH Terms
Descriptor/Qualifier:
Cell Differentiation / drug effects*
Cell Division / drug effects
Cell Survival
Cholecalciferol / pharmacology*
Culture Media
HL-60 Cells* / cytology
Humans
Monocytes / cytology
Neutrophils / cytology
Tretinoin / pharmacology*
Chemical
Reg. No./Substance:
0/Culture Media; 302-79-4/Tretinoin; 67-97-0/Cholecalciferol

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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