Isolation of planarian X-ray-sensitive stem cells by fluorescence-activated cell sorting
Article first published online: 13 JUL 2006
Development, Growth & Differentiation
Volume 48, Issue 6, pages 371–380, August 2006
How to Cite
Hayashi, T., Asami, M., Higuchi, S., Shibata, N. and Agata, K. (2006), Isolation of planarian X-ray-sensitive stem cells by fluorescence-activated cell sorting. Development, Growth & Differentiation, 48: 371–380. doi: 10.1111/j.1440-169X.2006.00876.x
- Issue published online: 13 JUL 2006
- Article first published online: 13 JUL 2006
- Received 6 April 2006; revised 12 May 2006; accepted 14 May 2006.
- stem cell;
- X-ray irradiation
The remarkable capability of planarian regeneration is mediated by a group of adult stem cells referred to as neoblasts. Although these cells possess many unique cytological characteristics (e.g. they are X-ray sensitive and contain chromatoid bodies), it has been difficult to isolate them after cell dissociation. This is one of the major reasons why planarian regenerative mechanisms have remained elusive for a long time. Here, we describe a new method to isolate the planarian adult stem cells as X-ray-sensitive cell populations by fluorescence-activated cell sorting (FACS). Dissociated cells from whole planarians were labeled with fluorescent dyes prior to fractionation by FACS. We compared the FACS profiles from X-ray-irradiated and non-irradiated planarians, and thereby found two cell fractions which contained X-ray-sensitive cells. These fractions, designated X1 and X2, were subjected to electron microscopic morphological analysis. We concluded that X-ray-sensitive cells in both fractions possessed typical stem cell morphology: an ovoid shape with a large nucleus and scant cytoplasm, and chromatoid bodies in the cytoplasm. This method of isolating X-ray-sensitive cells using FACS may provide a key tool for advancing our understanding of the stem cell system in planarians.