Document Detail


Selective dissociation and characterization of cells from different regions of multicell tumor spheroids.
MedLine Citation:
PMID:  7471046     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A technique has been developed which takes advantage of the spherical symmetry of EMT6/Ro multicellular tumor spheroids to isolate subpopulations of cells originating from various locations in the spheroid structure. The method involves gently exposing spheroids to a dilute trypsin solution at 18 to 20 degrees in specially designed dishes on a rotary shaker. Under these conditions, cells are released only from the outer spheroid surface; successive trypsin treatments dissociate cells from increasing depths in the spheroid. Measurements of the spheroid diameter and rate of cell dissociation demonstrated the reproducibility of the technique. Observations of histological sections showed that trypsin was active only over a small distance into the spheroid. Characterization of the cells isolated indicated that cell volume, membrane integrity, and clonogenic capacity all decreased for cells located in inner spheroid regions. Autoradiography and DNA content analysis by flow cytometry established that outer region cells were actively proliferating while inner region cells were in a nonproliferative state. There was a significant number of cells arrested with S- and G2-phase DNA contents as well as a large number arrested with a G1 DNA content, similar to recent findings for nonproliferating cells in tumors.
Authors:
J P Freyer; R M Sutherland
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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:  Cancer research     Volume:  40     ISSN:  0008-5472     ISO Abbreviation:  Cancer Res.     Publication Date:  1980 Nov 
Date Detail:
Created Date:  1981-05-26     Completed Date:  1981-05-26     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2984705R     Medline TA:  Cancer Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  3956-65     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Aggregation
Cell Cycle
Cell Division
Cell Survival
Cells, Cultured
Clone Cells / pathology
Female
Mammary Neoplasms, Experimental / pathology
Mice
Neoplasms, Experimental / pathology*
Grant Support
ID/Acronym/Agency:
CA-11051/CA/NCI NIH HHS; CA-11198/CA/NCI NIH HHS; CA-20329/CA/NCI NIH HHS

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


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