Abstract

Hematopoietic stem and progenitor cell (HSPC) transplantation is the paradigm for stem cell therapies. The protocol described here enables quantitative assessment of the body-wide HSPC reconstitution of different mature hematopoietic cells in mice based on their presence in circulating blood. The method determines donor-derived mature cell populations per mouse, over time, by quantitatively obtaining their absolute numbers in the peripheral blood and utilizing previously assessed tissue-distribution factors. A Markov-based birth/death computational model accounts for the drastic differences in mature cell half-lives. By quantifying the number of cells produced and eliminating host variability, the protocol can be used to directly compare the lineage output of different types of HSPCs on a per cell basis, thereby clarifying the lineage potential and expansion capacity of different cell populations. These protocols were developed for hematopoiesis, but can readily be extended to other contexts by simply replacing the cell types and distributions. HighlightsO_LIQuantitative assessment of stem and progenitor cell reconstitution capacity C_LIO_LIElimination of cell-specific recipient variability for accurate donor cell potential C_LIO_LIDirectly comparable lineage output within and between stem and progenitor cells C_LIO_LIBlood-based absolute quantification of whole-body repopulation over time C_LIO_LIMarkov modelling-based consideration of differential mature cell half-lives C_LI