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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.
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Stem Cell Research & Therapy

ISSN: 1757-6512