Figure 2.
Human lung–derived hematopoietic progenitors have in vivo engraftment potential. (A) Experimental procedure to compare the in vivo engraftment efficiency of lung and BM hematopoietic progenitors: after sublethal irradiation, NSG-SGM3 mice were injected IV with either 1.5 × 106 Live/Lin− human lung or BM cells. Ten weeks after transplantation, the BM, PB, and lung of recipient mice were collected and investigated for human cell engraftment. The graft properties are summarized in the table below. (B) Representative flow plots of human myeloid (hCD45++, hCD33+) and lymphoid (hCD45++, hCD19+) cell engraftment in the BM of a recipient mouse (+, upper panel) and nontransplanted control (−, lower panel). (C) Engraftment efficiency of human cells after xenotransplantation measured by flow cytometry. Bar graphs represent the percentage of hCD45++ cell engraftment in the BM, lung, and PB of recipient mice after transplantation of HSPCs from human BM (black) or lung (white). Mean ± SD; Student t test values are given, and individual data points for each animal are plotted as gray dots. Blue dotted line indicates threshold for positive engraftment. (D) Detection of human cells in the BM (left panel) and lung (right panel) of recipient mice by immunostaining against human CD45 (hCD45). Scale bar, 50 μm. (E) Representative flow cytometry plots of human erythroid (CD45−, hGlyA+, or hCD71+) cell engraftment in the BM of a recipient mouse (+, upper panel) and nontransplanted control (−, lower panel). (F) Human erythroid cell expansion (CD45−, hGlyA+, or hCD71+) in the BM, lung, and PB of recipient mice measured by flow cytometry. Bar graphs representing the percentage of CD45−hGlyA+hCD71+ cells in BM, lung, and PB after transplantation of HSPCs from human BM (black) or lung (white). Mean ± SD; Student t test values are given, and individual data points for each animal are plotted as gray dots. Blue dotted line indicates threshold for positive engraftment. (G) Detection of human erythroid cells in the BM (left panel) and lung (right panel) of recipient mice by immunostaining against human GlyA (hGlyA). Scale bar, 50 μm. (H) Proportion of lineage expansion across all human cells detected in the BM, lung, and PB, respectively. Stacked bars represent mean proportion ± SD; ANOVA followed by Sidak multiple comparison test; ns, not significant.

Human lung–derived hematopoietic progenitors have in vivo engraftment potential. (A) Experimental procedure to compare the in vivo engraftment efficiency of lung and BM hematopoietic progenitors: after sublethal irradiation, NSG-SGM3 mice were injected IV with either 1.5 × 106 Live/Lin− human lung or BM cells. Ten weeks after transplantation, the BM, PB, and lung of recipient mice were collected and investigated for human cell engraftment. The graft properties are summarized in the table below. (B) Representative flow plots of human myeloid (hCD45++, hCD33+) and lymphoid (hCD45++, hCD19+) cell engraftment in the BM of a recipient mouse (+, upper panel) and nontransplanted control (−, lower panel). (C) Engraftment efficiency of human cells after xenotransplantation measured by flow cytometry. Bar graphs represent the percentage of hCD45++ cell engraftment in the BM, lung, and PB of recipient mice after transplantation of HSPCs from human BM (black) or lung (white). Mean ± SD; Student t test values are given, and individual data points for each animal are plotted as gray dots. Blue dotted line indicates threshold for positive engraftment. (D) Detection of human cells in the BM (left panel) and lung (right panel) of recipient mice by immunostaining against human CD45 (hCD45). Scale bar, 50 μm. (E) Representative flow cytometry plots of human erythroid (CD45−, hGlyA+, or hCD71+) cell engraftment in the BM of a recipient mouse (+, upper panel) and nontransplanted control (−, lower panel). (F) Human erythroid cell expansion (CD45−, hGlyA+, or hCD71+) in the BM, lung, and PB of recipient mice measured by flow cytometry. Bar graphs representing the percentage of CD45−hGlyA+hCD71+ cells in BM, lung, and PB after transplantation of HSPCs from human BM (black) or lung (white). Mean ± SD; Student t test values are given, and individual data points for each animal are plotted as gray dots. Blue dotted line indicates threshold for positive engraftment. (G) Detection of human erythroid cells in the BM (left panel) and lung (right panel) of recipient mice by immunostaining against human GlyA (hGlyA). Scale bar, 50 μm. (H) Proportion of lineage expansion across all human cells detected in the BM, lung, and PB, respectively. Stacked bars represent mean proportion ± SD; ANOVA followed by Sidak multiple comparison test; ns, not significant.

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