These authors contributed equally to this work.
Single-cell gene profiling of planarian stem cells using fluorescent activated cell sorting and its “index sorting” function for stem cell research
Article first published online: 11 JAN 2010
© 2010 The Authors. Journal compilation © 2010 Japanese Society of Developmental Biologists
Development, Growth & Differentiation
Special Issue: Comparative Aspects of Stem Cells
Volume 52, Issue 1, pages 131–144, January 2010
How to Cite
Hayashi, T., Shibata, N., Okumura, R., Kudome, T., Nishimura, O., Tarui, H. and Agata, K. (2010), Single-cell gene profiling of planarian stem cells using fluorescent activated cell sorting and its “index sorting” function for stem cell research. Development, Growth & Differentiation, 52: 131–144. doi: 10.1111/j.1440-169X.2009.01157.x
- Issue published online: 11 JAN 2010
- Article first published online: 11 JAN 2010
- Received 22 October 2009; revised 16 November 2009; accepted 16 November 2009.
- fluorescent activated cell sorting-based single cell polymerase chain reaction;
- index sorting;
- stem cell
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.