Document Detail

Differentiation of A31T6 proadipocytes to adipocytes: a flow cytometric analysis.
MedLine Citation:
PMID:  1735459     Owner:  NLM     Status:  MEDLINE    
A flow cytometric assay has been developed which provides precise, quantitative information on the accumulation of cytoplasmic triglycerides in individual A31T6 proadipocytes as they differentiate into adipocytes. The opportunity to measure multiple optical parameters on a cell-by-cell basis has enabled us to monitor phenotypic aspects of differentiation with a greater level of sensitivity than was previously possible. Using the fluorescent hydrophobic probe, Nile red, we have found that as a cell proceeds along the differentiation pathway, the gold fluorescence signal from the cell increases, reflecting the accumulation of cytoplasmic lipid droplets. The value of the ratio of gold to red fluorescence defines a cell as being differentiated or undifferentiated. The greater resolution afforded by this cytometric method over more conventional approaches has allowed us to determine (1) the presence of an undifferentiated population of cells whose existence is not detected by conventional phase microscopy, (2) that insulin is not required to drive differentiation in this system, though the rate of differentiation is increased when the cells are exposed to insulin in combination with dexamethasone, (3) that exposure to a combination of insulin and dexamethasone results in a lower accumulation of lipid in a cell than does exposure to either agent alone, and (4) that A31T6 cells show the same response to differentiation-promoting agents whether applied at the time of plating or at confluence.
M J Smyth; W Wharton
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Experimental cell research     Volume:  199     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  1992 Mar 
Date Detail:
Created Date:  1992-03-10     Completed Date:  1992-03-10     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  29-38     Citation Subset:  IM    
Cellular and Molecular Biology Group, Life Sciences Division, Los Alamos National Laboratory, New Mexico 87545.
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MeSH Terms
Adipose Tissue / chemistry,  cytology*,  drug effects
Cell Differentiation / drug effects,  physiology
Cell Line
Cytoplasm / chemistry*
Dexamethasone / pharmacology
Flow Cytometry
Insulin / pharmacology
Microscopy, Fluorescence
Triglycerides / analysis*
Grant Support
Reg. No./Substance:
0/Oxazines; 0/Triglycerides; 11061-68-0/Insulin; 50-02-2/Dexamethasone; 7385-67-3/nile red

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