Document Detail


Establishment and optimization of a flow cytometric method for evaluation of viability of CD34+ cells after cryopreservation and comparison with trypan blue exclusion staining.
MedLine Citation:
PMID:  15987368     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Trypan blue exclusion staining is probably the most frequently applied method (Method I) for assessment of viability in peripheral blood progenitor cell grafts after cryopreservation. Alternatively, a flow cytometry-based method (Method II) was established and optimized. STUDY DESIGN AND METHODS: In a first series of 22 autologous apheresis products, the influence of duration of antibody staining and red cell (RBC) lysis on viability was investigated. In a second series of 21 autologous and 1 allogeneic apheresis products, the effect of omitting the RBC lysis was evaluated. On the basis of the results of the first two series, 155 autologous and 57 allogeneic apheresis products were analyzed with Method I and the now optimized Method II. RESULTS: Halving the incubation times did not influence the viability of CD45+ or CD34+ cells. Omission of RBC lysis resulted in a significantly (p = 0.022) increased median viability of CD45+ cells (75.8% vs. 71.0%) without any influence on CD34+ cells. In the third series, the median viability of CD34+ cells (96.9%) was significantly (p < 0.0001) higher compared with the viability of CD45+ cells (76.2%) and the viability determined by Method I (86.5%). CONCLUSION: The viability of CD34+ cells was significantly higher compared with the viability of all white blood cells. The presented cytometry-based method is superior to the standard trypan blue method regarding the number of analyzable cells and documentation, regarding observer independence and standardization; it allows the analysis of the cells of interest for transplantation after minimal sample manipulation.
Authors:
Andreas Humpe; Christian Beck; Robert Schoch; Michael Kneba; Heinz-August Horst
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article    
Journal Detail:
Title:  Transfusion     Volume:  45     ISSN:  0041-1132     ISO Abbreviation:  Transfusion     Publication Date:  2005 Jul 
Date Detail:
Created Date:  2005-06-30     Completed Date:  2005-08-25     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0417360     Medline TA:  Transfusion     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1208-13     Citation Subset:  IM    
Affiliation:
Second Department of Medicine of the University Clinic Schleswig-Holstein Campus Kiel, Dr Mildred Scheel Haus, Kiel, Germany. a.humpe@med2.uni-kiel.de
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MeSH Terms
Descriptor/Qualifier:
Antigens, CD34 / metabolism*
Antigens, CD45 / metabolism
Blood Component Removal
Cell Survival
Coloring Agents
Cryopreservation*
Evaluation Studies as Topic
Flow Cytometry*
Hemolysis
Humans
Staining and Labeling / methods*
Trypan Blue / metabolism*
Chemical
Reg. No./Substance:
0/Antigens, CD34; 0/Coloring Agents; 72-57-1/Trypan Blue; EC 3.1.3.48/Antigens, CD45

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


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