Figure 1.
Fluorescence-activated cell sorting of lineage negative (FITC −) leukocytes to isolate long term- and short term-hematopoietic stem cell groups. A) Forward and side scatter plot isolates cells. B) Forward scatter and PI plot captures living cells (PI−). C) Forward scatter and FITC plot removes remaining lineage+ cells (FITC+) and retains lineage− cells (FITC −). D) Sca-1 (Alexa Fluor 405+) and c-kit (APC +) positive cells are isolated. E) Forward scatter and Flk-2 plot separates long term cells (Flk-2− PE−) from short term cells (Flk-2+ PE+). Stem cells in the LT-HSC (Flk-2−) (F1, F2) and ST-HSC (Flk-2+) (G1, G2) groups were resorted to ensure that contaminating cells are not present in either population.
Figure 2.
Experiment 1: intravenous injection.
Chimerism (%CD45.1) (ordinate) plotted as a function of Days Post Transplant (abscissa), type of cells injected (•▪: LT-HSC, ○□: ST-HSC), and source (•○: whole blood, ▪□: whole bone marrow) for several different methods of summary: A) raw individual mouse percent CD45 cells that were CD45.1, B) mean ± standard error of the mean, C) median with inter-quartile range, and D) percent of mice with greater than 10% of CD45 cells being CD45.1. For each tile, the data from the primary transplant is plotted on the left and secondary transplant on the right, separated by a vertical line at zero for the secondary transplant.
Figure 3.
Experiment 1: intraperitoneal injection.
Chimerism (%CD45.1) (ordinate) plotted as a function of Days Post Transplant (abscissa), type of cells injected (•▪: LT-HSC, ○□: ST-HSC), and source (•○: whole blood, ▪□: whole bone marrow) for several different methods of summary: A) raw individual mouse percent CD45 cells that were CD45.1, B) mean ± standard error of the mean, C) median with inter-quartile range, and D) percent of mice with greater than 10% of CD45 cells being CD45.1. For each tile, the data from the primary transplant is plotted on the left and secondary transplant on the right, separated by a vertical line at zero for the secondary transplant.
Figure 4.
Experiment 2: intravenous injection.
Chimerism (%CD45.1) (ordinate) plotted as a function of Days Post Transplant (abscissa), type of cells injected (•▪: LT-HSC, ○□: ST-HSC), and source (•○: whole blood, ▪□: whole bone marrow) for several different methods of summary: A) raw individual mouse percent CD45 cells that were CD45.1, B) mean ± standard error of the mean, C) median with inter-quartile range, and D) percent of mice with greater than 10% of CD45 cells being CD45.1. For each tile, the data from the primary transplant is plotted on the left and secondary transplant on the right, separated by a vertical line at zero for the secondary transplant.
Figure 5.
Experiment 2: intrafemoral injection.
Chimerism (%CD45.1) (ordinate) plotted as a function of Days Post Transplant (abscissa), type of cells injected (•▪: LT-HSC, ○□: ST-HSC), and source (•○: whole blood, ▪□: whole bone marrow) for several different methods of summary: A) raw individual mouse percent CD45 cells that were CD45.1, B) mean ± standard error of the mean, C) median with inter-quartile range, and D) percent of mice with greater than 10% of CD45 cells being CD45.1. For each tile, the data from the primary transplant is plotted on the left and secondary transplant on the right, separated by a vertical line at zero for the secondary transplant.
Figure 6.
Experiment 3: intravenous injection.
Chimerism (%CD45.1) (ordinate) plotted as a function of Days Post Transplant (abscissa), type of cells injected (•▪: LT-HSC, ○□: ST-HSC), and source (•○: whole blood, ▪□: whole bone marrow) for several different methods of summary: A) raw individual mouse percent CD45 cells that were CD45.1, B) mean ± standard error of the mean, C) median with inter-quartile range, and D) percent of mice with greater than 10% of CD45 cells being CD45.1. For each tile, the data from the primary transplant is plotted on the left and secondary transplant on the right, separated by a vertical line at zero for the secondary transplant.
Figure 7.
Experiment 3: intrafemoral injection.
Chimerism (%CD45.1) (ordinate) plotted as a function of Days Post Transplant (abscissa), type of cells injected (•▪: LT-HSC, ○□: ST-HSC), and source (•○: whole blood, ▪□: whole bone marrow) for several different methods of summary: A) raw individual mouse percent CD45 cells that were CD45.1, B) mean ± standard error of the mean, C) median with inter-quartile range, and D) percent of mice with greater than 10% of CD45 cells being CD45.1. For each tile, the data from the primary transplant is plotted on the left and secondary transplant on the right, separated by a vertical line at zero for the secondary transplant.
Figure 8.
Multilineage nature of engraftment: Summary graph of all primary transplant experiments showing that when engraftment occurred, multiple cell types were seen.
Therefore, points displayed represent percent of cells from donor. Serial transplants of the primary whole bone marrow were also multilineage in nature when engraftment occurred (data not shown).