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Adaptation to ex vivo culture drives human haematopoietic stem cell loss of repopulation capacity in a cell cycle independent manner

Johnson, C. S.; Sham, K.; Belluschi, S.; Wang, X.; Lau, W.; Kaufmann, K. B.; Krivdova, G.; Calderbank, E. F.; Mende, N.; McLeod, J.; Mantica, G.; Williams, M. J.; Grey-Wilson, C.; Drakopoulos, M.; Sinha, S.; Diamanti, E.; Basford, C.; Green, A. R.; Wilson, N. K.; Howe, S. J.; Dick, J. E.; Gottgens, B.; Francis, N.; Laurenti, E.

2022-11-17 cell biology
10.1101/2022.11.17.516906 bioRxiv
Show abstract

Loss of long-term haematopoietic stem cell function (LT-HSC) hampers the success of ex vivo HSC gene therapy and expansion procedures, but the kinetics and the mechanisms by which this occurs remain incompletely characterized. Here through time-resolved scRNA-Seq, matched in vivo functional analysis and the use of a reversible in vitro system of early G1 arrest, we define the sequence of transcriptional and functional events occurring during the first ex vivo division of human LT-HSCs. We demonstrate that contrary to current assumptions, loss of long-term repopulation capacity during culture is independent of cell cycle progression. Instead it is a rapid event that follows an early period of adaptation to culture, characterised by transient gene expression dynamics and constrained global variability in gene expression. Cell cycle progression however contributes to the establishment of differentiation programmes in culture. Our data have important implications for improving HSC gene therapy and expansion protocols.

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