Document Detail


Single-cell gene profiling of planarian stem cells using fluorescent activated cell sorting and its "index sorting" function for stem cell research.
MedLine Citation:
PMID:  20078655     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To achieve an integrated understanding of the stem cell system of planarians at both the cellular and molecular levels, we developed a new method by combining "fluorescent activated cell sorting (FACS) index sorting" analysis and single-cell reverse transcription-polymerase chain reaction (RT-PCR) to detect the gene expression and cell cycle state of stem cells simultaneously. Single cells were collected using FACS, and cDNAs of each cell were used for semi-quantitative RT-PCR. The results were plotted on the FACS sorting profile using the "index sorting" function, which enabled us to analyze the gene expression in combination with cell biological data (such as cell cycle phase) for each cell. Here we investigated the adult stem cells of planarians using this method and obtained findings suggesting that the stem cells might undergo commitment during S to G2/M phase. This method could be a powerful and straightforward tool for examining the stem cell biology of not only planarians but also other organisms, including vertebrates.
Authors:
Tetsutaro Hayashi; Norito Shibata; Ryo Okumura; Tomomi Kudome; Osamu Nishimura; Hiroshi Tarui; Kiyokazu Agata
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Development, growth & differentiation     Volume:  52     ISSN:  1440-169X     ISO Abbreviation:  Dev. Growth Differ.     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-01-18     Completed Date:  2010-04-27     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0356504     Medline TA:  Dev Growth Differ     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  131-44     Citation Subset:  IM    
Affiliation:
RIKEN Center for Developmental Biology, Chuo-ku, Kobe 650-0047, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Division / genetics
Cell Separation / methods
Cluster Analysis
Female
Flow Cytometry / methods
G2 Phase / genetics
Gene Expression Profiling*
Genes, Helminth / genetics
Helminth Proteins / genetics
In Situ Hybridization
Male
Planarians / cytology*,  radiation effects
Proliferating Cell Nuclear Antigen / genetics
Research / methods
Reverse Transcriptase Polymerase Chain Reaction
S Phase / genetics
Stem Cells / cytology*,  metabolism*
Chemical
Reg. No./Substance:
0/Helminth Proteins; 0/Proliferating Cell Nuclear Antigen

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