Looking for the elusive lung stem cell niche

Translational Respiratory Medicine

Table 2 Number of hematopoietic and non-hematopoietic cells in BALf and lung a week after bleomycin treatment- Trends in cellular migration to BALf and Lung post-bleomycin overtime

	WT saline (d7)	WT bleo (d7)	NOX saline (d7)	NOX bleo (d7)	DRO saline (d7)	DKO bleo (d7)
Totalcellcontent (xlIYIml)	4.27 ± 0.184	28.3 ± 0224	4.515 ± 0.223	48.91* ± 1.59	2.61 ± 0.16	8.42 ± 0.13
Macrophaqes (xlIYIml)	4.16 ± 0.57 (972%)	11.14 ± 4.08 (37,6%)	4.01 ± 0.31 (89.9%)	28.14* ± 6.83 (57.47%)	1.87 ± 0.64 (71.64%)	4.19 ± 3.18 (49.82%)
Lymph ocytes (xWlml)	0.08 ± 0.01 (2.0%)	12.63 ± 2.49 (46.0%)	0.09 ± 0.02 (2.01%)	16.91 3.65ç34.53%)	1.2 ± 0.01 (4.59%)	1 .44 0.66 (17.12%)
Neutrophils (x’llYIml)	0.03 ± 0.01 (0.7%)	4.5 ± 0.97 (16.3%)	0.11 ± 0.01 (0.02%)	3.91 ± 053 (7.98%)	0.63 ± 0.14 (24.13 96)	278 ± 1.06 (33%)

Number of cells in BALf after a week of bleomycin treatment.
After the single intra-tracheal dose of bleomycin, animals were sacrificed on d7, d14, and d21 for the WT after bleo, and at D7 and d21 only for NOX-/- and DKO mice and wildtype saline treated mice. Cell number was quantitated by hemocytometer and corroborated by Z1 particle counter (Beckman Coulter). Number of cells represent data from 2 independent experiments ± SEM. N = 4/group. Cell subsets were quantified by FACS and morphologic differentiation under light microscope. Olympus BX41 at 40× magnification. Abbreviations used are: WT (wild type), NOX-/- (gp91phox knockout), gp91phox MMp12 double knockout (DKO).CD45+ cells were gated and macrophages (Gr1-F4/80hi), lymphocytes (CD3+ and B220+), and neutrophils (Gr-1hi F4/80-) were identified, their percentages analyzed by CellQuestPro and their total numbers calculated from the total number of cells obtained by lavage as described in Materials and methods. Data presented is pooled from 2 independent experiments ± SEM. *denotes p value < 0.05 compared to WT post-bleo on corresponding time points. Abbreviations used are: WS = Wildtype (WT) + saline (d7), WB7 = WT + post-bleomycin d7, WB14 = WT + post-bleomycin d14, WB21 = WT + post-bleomycin d21, NOXS = gp91phox-/- + saline d21, NOXB21 = gp91phox-/- + bleomycin d21, DKOS = gp91phox-MMP12 double knockout + saline (d21), DKOB21 = gp91phox-MMP12 double knockout + bleomycin (d21).