Document Detail


Flow cytometric evaluation of cell dispersion from human head and neck tumors.
MedLine Citation:
PMID:  2990834     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The preparation of single-cell suspensions from 25 human head and neck tumors is described. Dispersal was performed overnight at 4 degrees C under slight agitation of the tissue suspensions using various combinations of enzymes and additives. The cell suspensions were examined for number of cells released, viability, amount of debris, and DNA distribution by means of flow cytometry (FCM). It was shown that both trypsin/dithioerythritol (TD) and collagenase/D Nase (CDse) were of value in dispersing single cells from tumor tissue. In contrast to CDse, incubation with TD appeared to be cytolytic to normal lymphocytes. In a number of cases, DNA-FCM revealed ploidy abnormalities in a TD-suspension, which were not discernible in the concurrent CDse-suspension. Cell culture of primary cell suspensions corroborated the reliability of the DNA-FCM measurements. Pretreatment with CDse improved tumor disaggregation by TD and indicated a different dispersal capacity. Addition of Ca2+ and Mg2+ ions to the dispersal mixtures and preincubation of tumor slices in complete medium for 1 day before initiation of cell dispersion influenced favorably the quality of the cell suspension.
Authors:
J T Bijman; D J Wagener; H van Rennes; J M Wessels; P van den Broek
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry     Volume:  6     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  1985 Jul 
Date Detail:
Created Date:  1985-09-23     Completed Date:  1985-09-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  334-41     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Adult
Aged
Cell Separation
Chlorides / diagnostic use
DNA, Neoplasm / analysis
Deoxyribonuclease I / diagnostic use
Dithioerythritol / diagnostic use
Female
Flow Cytometry
Head and Neck Neoplasms / analysis,  pathology*
Humans
Hyaluronoglucosaminidase / diagnostic use
Lithium / diagnostic use
Lithium Chloride
Male
Microbial Collagenase / diagnostic use
Middle Aged
Pepsin A / diagnostic use
Ribonuclease, Pancreatic / diagnostic use
Trypsin / diagnostic use
Chemical
Reg. No./Substance:
0/Chlorides; 0/DNA, Neoplasm; 6892-68-8/Dithioerythritol; 7439-93-2/Lithium; 7447-41-8/Lithium Chloride; EC 3.1.21.1/Deoxyribonuclease I; EC 3.1.27.5/Ribonuclease, Pancreatic; EC 3.2.1.35/Hyaluronoglucosaminidase; EC 3.4.21.4/Trypsin; EC 3.4.23.1/Pepsin A; EC 3.4.24.3/Microbial Collagenase

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


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