Early hemopoietic progenitor cells: direct measurement of cell cycle status

We have investigated the cell cycle status of murine hemopoietic progenitors using vital DNA staining and flow sorting. Suspended Balb/c bone marrow cells were stained with Hoechst 33342 dye and separated first on light scattering properties; this procedure allowed a 5-fold enrichment in progenitor cells. A second sorting based on DNA content indicated that 80% of these cells were in G0/G1 and 20% in S-G2 + M. When G0/G1 and S-G2 + M cells were assayed separately in methylcellulose cultures, or with the in vivo colony forming assay, the G0/G1 cells were shown to be markedly enriched in CFU-S, BFU-E, and GM-CFU as compared to S-G2 + M cells with the final recovery increased 20-fold. Comparison of different strains or age groups yielded results identical to those obtained with Balb/c with the exception of C57B1/6. In the latter strain only a 3-fold enrichment could be observed in the G0/G1 fraction. These results demonstrate that the majority of early hemopoietic progenitors are in the G0/G1 phase of the cell cycle.