Document Detail

"Microenvironmental contaminations" induced by fluorescent lipophilic dyes used for noninvasive in vitro and in vivo cell tracking.
MedLine Citation:
PMID:  20215639     Owner:  NLM     Status:  MEDLINE    
Determining how normal and leukemic stem cells behave in vivo, in a dynamic and noninvasive way, remains a major challenge. Most optical tracking technologies rely on the use of fluorescent or bioluminescent reporter genes, which need to be stably expressed in the cells of interest. Because gene transfer in primary leukemia samples represents a major risk to impair their capability to engraft in a xenogenic context, we evaluated the possibility to use gene transfer-free labeling technologies. The lipophilic dye 3,3,3',3' tetramethylindotricarbocyanine iodide (DiR) was selected among 4 near-infrared (NIR) staining technologies. Unfortunately we report here a massive transfer of the dye occurring toward the neighbor cells both in vivo and in vitro. We further demonstrate that all lipophilic dyes tested in this study (1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine perchlorate [DiI], DiD, DiR, and PKH26) can give rise to microenvironmental contamination, including when used in suboptimal concentration, after extensive washing procedures and in the absence of phagocytosis or marked cell death. This was observed from all cell types tested. Eventually, we show that this microenvironmental contamination is mediated by both direct cell-cell contacts and diffusible microparticles. We conclude that tracking of labeled cells using non-genetically encoded markers should always be accompanied by drastic cross validation using multimodality approaches.
Francois Lassailly; Emmanuel Griessinger; Dominique Bonnet
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-03-09
Journal Detail:
Title:  Blood     Volume:  115     ISSN:  1528-0020     ISO Abbreviation:  Blood     Publication Date:  2010 Jul 
Date Detail:
Created Date:  2010-07-02     Completed Date:  2010-08-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7603509     Medline TA:  Blood     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5347-54     Citation Subset:  AIM; IM    
Hematopoietic Stem Cell Laboratory, London Research Institute, Cancer Research UK, 44 Lincoln's Inn Fields, London, United Kingdom.
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MeSH Terms
Cell Line
Cells, Cultured
Coculture Techniques
Fetal Blood / cytology
Flow Cytometry* / methods
Fluorescent Dyes / metabolism*
Hematopoietic Stem Cells / cytology
Microscopy, Confocal
Spectroscopy, Near-Infrared / methods
Whole Body Imaging* / methods
Grant Support
//Cancer Research UK
Reg. No./Substance:
0/Fluorescent Dyes

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