1. Engraftment of Bone Marrow–derived Stem Cells to the Lung in a Model of Acute Respiratory Infection by Pseudomonas aeruginosa 
Joanna Rejman, Carla Colombo, Massimo Conese 
Molecular Therapy 
Volume 17, Issue 7, Pages (July 2009)
DOI: /mt
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Epithelial integrity and vascular permeabilty in the airways of mice infected with Pseudomonas aeruginosa. (a) Total protein, (b) albumin, and (c) lactate dehydrogenase (LDH) levels in bronchioalveolar lavage fluid (BALF) of control and treated animals. C57Bl/6 mice were inoculated with 105 or 106 cfu of P. aeruginosa. The mice were killed 24 (white bars) or 48 hours (black bars) postinstillation and their BALF was recovered. The graphs represent means ± SD. *P < 0.05 (versus control value); n = 5. cfu, colony forming units.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
Histological assessment of respiratory epithelial damage in the airways following infection with Pseudomonas aeruginosa. (a) Hematoxylin and eosin-stained lung section from control mice and (b,c) hematoxylin and eosin-stained lung sections from P. aeruginosa-infected mice. Bacteria injected at a dose of 105 cfu induced only local damage to the respiratory epithelium (b). Evidence of extensive damage can be observed on lung sections from mice inoculated with P. aeruginosa at a dose of 106 cfu (c). The mice were killed 48 hours postinstillation. Micrographs are representative of lung sections obtained from three animals per group. Staining was performed on 5–10 sections per lobe. Original Magnification ×20. cfu, colony forming units.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
Detection of donor-derived GFP+ cells in mouse lung sections. Recipient C57Bl/6 mice were inoculated with Pseudomonas aeruginosa at a dose of 106 cfu. 100 µl of PBS was instilled into the trachea of control mice. Adult transgenic C57Bl/6 mice constitutively expressing GFP were used as donors. Bone marrow cells isolated from these mice were enriched in progenitor cells by means of the EasySep system. The cell suspension was injected intratracheally in mice infected with P. aeruginosa 2 days earlier. The mice were killed after 6 weeks and the lung sections were screened for GFP+ cells. The presence of donor-derived GFP+ cells was evaluated by immunochemistry with an anti-GFP antibody and using avidin-biotin-peroxidase detection system. GFP-expressing cells were readily detected in the lungs of the surviving mice inoculated with P. aeruginosa at a dose of 106 cfu. (a) Control mice (PBS + GFP) and (b) treated mice (bacteria + GFP). Arrows indicate donor-derived GFP+ cells. Micrographs are representative of lung sections obtained from three animals per group. Staining was performed on 10–15 sections per lobe. Original Magnification ×40. cfu, colony forming units; GFP, green fluorescent protein; PBS, phosphate buffered saline.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
Donor-derived GFP+ cells are located in the bronchial/bronchiolar lining and express cytokeratins (CKs). (a,b) Presence of donor-derived GFP+/CK+ cells evaluated by direct GFP fluorescence and immunofluorescence in Pseudomonas aeruginosa-infected mice. Recipient C57Bl/6 mice were inoculated with P. aeruginosa at a dose of 106 cfu. Adult transgenic mice C57Bl/6 mice constitutively expressing GFP were used as donors. Bone marrow cells isolated from these mice were enriched in progenitor cells by means of the EasySep system. The cell suspension was injected intratracheally in mice infected with P. aeruginosa 2 days earlier. The mice were killed after 6 weeks and the lung sections were screened for GFP+ cells and CK+ cells. Donor-derived GFP+ cells (green) located in the bronchial lining expressed cytokeratins (red), which indicates that these cells are of an epithelial nature. Nuclei are stained with DAPI. Micrographs are representative of lung sections obtained from three animals per group. Staining was performed on 8–15 sections per lobe. Original magnification ×40. cfu, colony forming units; DAPI, 4′-6-Diamidino-2-phenylindole; GFP, green fluorescent protein.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

6. Figure 5
Donor-derived Y-chromosome+ cells are embedded in the basal and columnar layers of the respiratory epithelium. Recipient female C57Bl/6 mice were inoculated with Pseudomonas aeruginosa at a dose of 106 cfu. Male C57Bl/6 mice were used as donors. Bone marrow cells isolated from these mice were enriched in progenitor cells using the EasySep system. The cell suspension was injected intratracheally in mice infected with P. aeruginosa 2 days earlier. The mice were killed after 6 weeks and the lung sections were screened for Y-chromosome+ cells and cytokeratin (CK+) cells. (a) Y-chromosome+ cells (green) that are CK+ (red). (b,c) Enlarged micrographs of image a. (d,f) Other examples of Y-chromosome+ cells that express CKs. Arrows indicate donor-derived CK+ cells. (g–i) Images were taken at a confocal laser scanning microscope; g and h show Y-chromosome+ cells, (i) Y-chromosome+ cells (green) that are CK+ (red). Note that donor-derived cells are present in the basal and columnar layers of the respiratory epithelium. Micrographs are representative of lung sections obtained from four animals. Original magnification ×63, except b, c, h, and i (×100). cfu, colony forming units.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions