Isolation of hematopoietic stem cells by single marker

by Alexey Bersenev on September 10, 2014 · 0 comments

in hematopoietic

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Hematopoietic stem cells (HSC) is the first type of adult stem cells, which was prospectively isolated by defined markers. Characterization of mouse and human HSCs and their isolation by FACS have gotten more and more sophisticated with years. Now, it typically includes combination of 6-9 surface and “functional” markers. But attempts to identify unique HSC marker remain elusive.

Recently, Derrick Rossi’s lab from Harvard University described unique single marker for mouse HSCFgd5. They started from mining the data for identification of unique gene expression in purified HSC, available from public datasets. They were able to narrow down search to 3 genes, restricted to HSCs. Then they generated reporter mice with fluorescent label for each of 3 genes. Only immunophenotypic HSCs from Fgd5 reporter mice were uniformly positive for fluorescent label mCherry. mCherry+ bone marrow cells from these reporter mice have immunophenotype of HSC and possess all HSC functions, tested in serial transplantation, limiting dilution and multilineage chimerism assays.

So, finally we got an unique HSC marker! The twist is that usability of this marker can be utilized only in special “genetically designed” mice. You cannot simply sort HSC by Fgd5 from wild type mice. This is not a surface marker.

Margaret Goodell highlighted the significance of this study for HSC research in commentary:

… establishing these Fdg5-reporter mouse lines and demonstrating the high and specific expression pattern that can be achieved with insertions into this locus is a significant contribution to the field. The power of the study lies in the utility of the mouse lines, but it also provides a benchmark for such reporters in the future.

This study demonstrates a way to identify unique markers for defined adult stem cells. Potential value for research right now tied to “Fgd5 reporter mouse line”. But I’m dreaming about commercial kit for HSC isolation by single marker from bone marrow cell suspension within an hour. Fgd5 could be a candidate for developing such kit. Maybe molecular beacons can do Fgd5 without affecting HSC functionality?

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