Local clonal analysis of the hematopoietic system shows that multiple small short-living clones maintain life-long hematopoiesis in reconstituted mice

Abstract

We describe here a technique to study the clonal contribution of primitive stem cells that account for long-term hematopoiesis in the same mouse over a 14-month period. Specifically, irradiated recipient female mice were transplanted with retrovirally marked male hematopoietic progenitors. Bone marrow was then collected repeatedly from local sites from the same mice throughout a 14-month period and injected into secondary irradiated recipients for analysis of donor retrovirally marked day-11 colony-forming unit-spleen (CFU-S-11). We have tracked the temporal in vivo fate of 194 individual CFU-S-derived cell clones in 38 mice reconstituted with such retrovirally marked bone marrow cells. Our data show that long-term hematopoiesis is maintained by a large number of simultaneously functioning small, shortlived (1 to 3 months) clones that usually grow locally with little or no dispersion between different regions of the hematopoietic system. Furthermore, the clones that disappeared were never detected again. The data suggest that normal hematopoiesis is supported by the sequential recruitment of marrow repopulating cells into a differentiation mode.

Stem Cell Defect in Ubiquitin-Green Fluorescent Protein Mice Facilitates Engraftment of Lymphoid-Primed Hematopoietic Stem Cells

“…In parallel with these various approaches enabling the distinction of donor and host blood cells, advancements in molecular biology enabled cells to be tagged with artificial a Institute of Pathological Physiology, First Faculty of Medicine, Charles University, genetic markers [13][14][15][16] including the current advanced technique of cell barcoding [17][18][19][20]. The insertion of artificial gene constructs expressing various types of fluorescent proteins also became an important research tool in experimental hematology and immunology as well as in other fields of the biomedical research [21,22].…”

Stable Long-Term Blood Formation by Stem Cells in Murine Steady-State Hematopoiesis

“…In a hierarchical structure, multipotent long‐term HSCs give rise to short‐term HSCs and lineage‐restricted progenitors that in turn differentiate to mature myeloid or lymphoid blood cells [1]. Early transplantation experiments using genetically marked hematopoietic cells revealed sequential fluctuations of murine short‐lived hematopoietic clones supporting a clonal succession model of stem cell activity [7–9]. In this model, at any given time, blood cell production is driven by a low number of HSC clones, whereas most other HSCs are quiescent but are recruited to replace exhausted clones [10].…”

The effects of proliferation and DNA damage on hematopoietic stem cell function determine aging

“…Importantly, this increased proliferation of HSCs led to depletion of their engraftment potential and overall function. This was concomitant with the observation that various subclones of hematopoietic progenitors exist in the adult BM with high proliferation rate, and they exhibit a significantly shorter life span (Drize et al, ). In fact, proliferation rate has been shown to be inversely proportional to the self‐renewal potential and overall stemness of HSCs.…”