A highly efficient reporter system for identifying and characterizing in vitro expanded hematopoietic stem cells

Che, J. L. C.; Bode, D.; Kucinski, I.; Cull, A. H.; Bain, F.; Barile, M.; Boyd, G.; Belmonte, M.; Jassinskaja, M.; Rubio-Lara, J.; Shepherd, M. S.; Clay, A.; Wilkinson, A. C.; Nakauchi, H.; Yamazaki, S.; Göttgens, B.; Kent, D. G.

doi:10.1101/2021.06.18.448972

bioRxiv · 10.1101/2021.06.18.448972

A highly efficient reporter system for identifying and characterizing in vitro expanded hematopoietic stem cells

Che, J. L. C.·Bode, D.·Kucinski, I.·Cull, A. H.·Bain, F.·Barile, M.·Boyd, G.·Belmonte, M.·Jassinskaja, M.·Rubio-Lara, J.·Shepherd, M. S.·Clay, A.·Wilkinson, A. C.·Nakauchi, H.·Yamazaki, S.·Göttgens, B.·Kent, D. G.

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Abstract

Hematopoietic stem cells (HSCs) cultured outside the body are the fundamental component of a wide range of cellular and gene therapies. Recent efforts have achieved more than 200-fold expansion of functional HSCs, but their molecular characterization has not been possible due to the substantial majority of cells being non-HSCs and single cell-initiated cultures displaying substantial clone-to-clone variability. Using the Fgd5 reporter mouse in combination with the EPCR surface marker, we report exclusive identification of HSCs from non-HSCs in expansion cultures. Linking single clone functional transplantation data with single clone gene expression profiling, we show that the molecular profile of expanded HSCs is similar to actively cycling fetal liver HSCs and shares a gene expression signature with functional HSCs from all sources, including Prdm16, Fstl1 and Palld. This new tool can now be applied to a wide-range of functional screening and molecular experiments previously not possible due to limited HSC numbers.

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Che, J. L. C., Bode, D., Kucinski, I., Cull, A. H., Bain, F., Barile, M., Boyd, G., Belmonte, M., Jassinskaja, M., Rubio-Lara, J., Shepherd, M. S., Clay, A., Wilkinson, A. C., Nakauchi, H., Yamazaki, S., Göttgens, B., Kent, D. G.. 2021-06-18. A highly efficient reporter system for identifying and characterizing in vitro expanded hematopoietic stem cells. https://doi.org/10.1101/2021.06.18.448972

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