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Table 2 Percent positive cells by FACS within BrdU+ and BrdU- populations in lung for pluripotent and pulmonary lineage-specific markers

From: Characterization of lung stem cell niches in a mouse model of bleomycin-induced fibrosis

A. Percent positive cells by FACS within BrdU+ population in lung
  BrdU+ TTF-1+ Oct3/4+ SSEA-3+ SSEA-4+ Sca-1+ Lin- CD34+ CD31+
Con 2d 8.51 ± 0.23 20.55 ± 0.30 86.18 ± 1.05 40.9 ± 0.83 48.85 ± 0.25 932 ± 1.70 95.54 ± 0.78 1.08 ± 0.02 2.39 ± 0.10
Con 6d 9.05 ± 0.03 21.11 ± 0.59 85.67 ± 0.93 42.90 ± 1.11 46.44 ± 2.24 87.66 ± 2.56 92.30 ± 0.66 1.58 ± 0.20 1.92 ± 0.63
Bleo 2d 2.67 ± 0.12 4.16 ± 0.12 78.25 ± 2.01 8.91 ± 0.02 8.97 ± 0.025 3.67 ± 0.09 44.82 ± 1.75 2.42 ± 0.17 4.77 ± 0.18
Bleo 4d 1.83 ± 0.21 4.52 ± 0.45 80.58 ± 0.12 8.07 ± 0.15 8.57 ± 0.41 3.71 ± 0.30 38.05 ± 1.54 3.01 ± 0.49 4.60 ± 0.19
Bleo 6d 1.87 ± 0.07 4.37 ± 0.29 79.62 ± 0.74 8.12 ± 0.53 7.95 ± 0.043 4.46 ± 0.24 36.28 ± 1.79 3.32 ± 0.40 3.60 ± 0.26
  SP-C+ AQP-5 CC-10 CD45+ CD45- SP-C+
CC10+
VEGF+
Con 2d 4.69 ± 0.13 0.83 ± 0.02 2.52 ± 0.15 2.13 ± 0.02 96.93 ± 0.90 82.80 ± 1.53 15.67 ± 0.86
Con 6d 4.59 ± 0.26 0.8 ± 0.004 2.42 ± 0.24 2.53 ± 0.23 93.95 ± 2.34 81.08 ± 0.42 13.78 ± 0.42
Bleo 2d 0.23 ± 0.05 0.14 ± 0.01 2.67 ± 0.09 3.42 ± 0.12 91.79 ± 1.60 2.43 ± 0.08 8.59 ± 0.29
Bleo 4d 0.54 ± 0.08 0.14 ± 0.02 3.16 ± 0.05 3.5 ± 0.20 90.38 ± 0.77 2.4 ± 0.56 8.70 ± 0.28
Bleo 6d 0.32 ± 0.04 0.14 ± 0.009 3.29 ± 0.12 3.57 ± 0.21 87.25 ± 2.29 2.08 ± 0.18 7.78 ±
B. Percent positive cells by FACS within BrdU - population in lung
  BrdU- TTF-
1+
Oct3/
4+
SSEA
-3+
SSEA
-4+
Sca-
1+
Lin- CD3
4+
CD3
1+
Con 2d 92.13 ± 0.84 0.38 ± 0.10 0.560 ± 0.09 0.55 ± 0.02 0.47 ± 0.14 0.22 ± 0.15 0.2 ± 0.01 0.31 ± 0.06 0.03 ± 0.01
Con 6d 92.95 ± 0.71 0.31 ± 0.07 0.35 ± 0.02 0.4 ± 0.07 0.32 ± 0.03 0.06 ± 0.02 0.19 ± 0.04 0.22 ± 0.009 0.22 ± 0.09
Bleo 2d 89.06 ± 0.66 11.36 ± 0.59 1.11 ± 0.09 0.28 ± 0.07 0.35 ± 0.02 0.12 ± 0.07 0.34 ± 0.03 3.45 ± 0.09 0.71 ± 0.042
Bleo 4d 89.43 ± 0.66 12.13 ± 0. 74 1.34 ± 0.09 0.32 ± 0.04 0.26 ± 0.08 0.25 ± 0.03 0.24 ± 0.08 3.42 ± 0.23 0.8 ± 0.09
Bleo 6d 97.40 ± 1.1 11.68 ± 0.6 1.742 ± 0.16 0.31 ± 0.04 0.32 ± 0.01 0.16 ± 0.05 0.54 ± 0.05 2.90 ± 0.06 0.6 ± 0.086
  SP-C+ AQP-5+ CC-10 CD45+ CD45- SP-C+
CC10+
VEGF+
Con 2d 5.59 ± 0.11 90.92 ± 0.50 1.30 ± 0.18 2.37 ± 0.17 80.38 ± 1.36 0.03 ± 0.01 2.86 ± 0.09
Con 6d 4.90 ± 0.14 85.75 ± 2.21 1.33 ± 0.21 2.7 ± 0.16 83.70 ± 1.66 0.12 ± 0.03 2.31 ± 0.14
Bleo 2d 1.48 ± 0.17 9.29 ± 0.41 2.60 ± 0.16 0.78 ± 0.06 76.15 ± 1.69 0.01 ± 0.004 12.74 ± 0.59
Bleo 4d 1.74 ± 0.03 217.49 ± 208.5 2.85 ± 0.08 0.65 ± 0.13 74.65 ± 1.37 0.02 ± 0.004 12.45 ± 0.84
Bleo 6d 1.52 ± 0.06 9.05 ± 0.18 2.36 ± 0.19 0.81 ± 0.13 74.05 ± 1.65 0.12 ± 0.06 10.65 ± 0.26
  1. Bromodeoxyuridine-positive (BrdU+) cells were gated and inside (positive, and outside of the gate, BrdU-negative) cells were characterized by FACS with fluorochrome conjugated monoclonal antibodies markers of pluripotence (TTF-1, SSEA-3,4, Oct ¾, SCA-1, lineage negative) and CD31 (endothelial progenitor) and CD34 (hematopoietic progenitor). Lung lineages are characterized by the following markers: AQP-5+ cells are AEI, SP-C+ cells are AEII and CC-10+ cells are Clara cells while SP-c+CC-10+ cells are bronchoalveolar stem cells. The gating strategies were as follows: BrdU+ and BrdU- cells were first gated and then the other markers were detected within the gates. Gating was first done on BrdU+ and BrdU- cells and then individual markers such as TTF-1, Oct3/4, SSEA 3, 4, SCA-1+, Lin-, CD34+ and CD31+ cells were assessed. The percentages, therefore, do not add up to 100% as only single positive cells are separately assessed in each gate. It is possible that there may be some redundancy but overall, it is possible have a rough idea about the kind of cells that are label-retaining cells and therefore the putative stem cells within the mouse lung. Con, control group; Bleo, bleomycin-treated group.