1. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 1. Characterization of MSC and alveolar A549 cells. (A) MSCs were harvested from human umbilical cord Wharton's jelly. MSC at passage 5 displayed a spindle-like shape, (B-D) MSC tri-lineage differentiation: adipogenesis, osteogenesis and chondrogenesis were confirmed by staining with Oil Red O, Alizarin Red and Alcian Blue, respectively. (Scale bars, 100 µm). (E) MSC was positive for mesenchymal stem cell markers CD105, CD73 and CD90 and negative for HLA-DR, CD11a, CD45 and CD34. (F-H) A human type II alveolar epithelial cell line, A549 was positive for type II AEC marker proSP-C. Phase contrast image is taken from an independent field to the proSP-C, DAPI and proSP-C/DAPI field. (Scale bars, 100 µm).
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

2. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 2. MSC-CM promotes proliferation and migration of alveolar epithelial cells (A549 cells). (A) Proliferation of A549 cells is increased by MSC-CM (50 ×) in a dose-dependent manner. There is no significant difference between 6 µL and 12 µL, suggesting 6 µL as an optimal dosage. Values given are means ±standard deviations of triplicate measurements. *Significant differences (P < 0.05) versus the LPS group are noted. (B) MSC-CM promotes the proliferation of A549 cells under septic conditions at day 1, 3 and 5 (*P < 0.05, ** P < 0.01). (C, E) In vitro wound healing assay. Scratches were made in the monolayer and cells were cultured for 24 hours in the absence or presence of MSC-CM. Representative areas are presented (Scale bar, 100 µm). (D, F) Transwell migration assay. Cells were treated with or without MSC-CM. After 16 hours of incubation, the cells at the upper side of the filter were mechanically removed. Cells that had migrated to the lower side of the filter were fixed for 10 minutes with 4% paraformaldehyde and stained using crystal violet for 1 hour. Ten random fields at a magnification of 200× were counted (Scale bar, 100µm). **Significant differences (P < 0.01) versus the LPS group are noted. *Significant differences (P < 0.05) versus the LPS group are noted.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

3. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 3. MSC-CM promotes proliferation and migration of primary human small airway epithelial cells (SAEC). (A) Proliferation of SAEC is increased by MSC-CM (50×) in a dose-dependent manner. There is no significant difference between 6 µL and 12 µL, suggesting 6 µL as an optimal dosage. Values given are means ±standard deviations of triplicate measurements. *Significant differences (P < 0.05) versus the LPS group are noted. (B) MSC-CM promotes the proliferation of SAEC under septic conditions at day 1 and 3 (** P < 0.01). (C, D) In vitro wound healing assay. Scratches were made in the monolayer and cells were cultured for 24 hours in the absence or presence of MSC-CM. Representative areas are presented (Scale bar, 100µm). ** Significant differences (P < 0.01) versus the LPS group are noted.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

4. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 4. MSC-CM activates JNK and P38 MAPK in A549 cells. (A-F) A549 cells were treated with or without MSC-CM for 0, 0.5, 1, 2, 6, or 12 hours after LPS stimulation. Cell lysates were prepared and used for Western blot analysis with phospho-JNK, total JNK (A-B), phospho-P38, total P38 (C-D), and phospho-ERK and total ERK (E-F). All experiments are representative of 3 replicates. ** Significant differences (P < 0.01) versus the LPS group are noted.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

5. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 5. MSC-CM activates JNK and P38 MAPK in primary human small airway epithelial cells (SAEC). (A-F) SAECs were treated with or without MSC-CM\for 0, 0.5, 1, 2, 6, or 12 hours after LPS stimulation. Cell lysates were prepared and used for Western blot analysis with phospho-JNK, total JNK (A-B), phospho-P38, total P38 (C-D), and phospho-ERK and total ERK (E-F). All experiments are representative of 3 replicates. ** Significant differences (P < 0.01) versus the LPS group are noted. *Significant differences (P < 0.05) versus the LPS group are noted.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

6. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 6. Blockage of the JNK and P38 MAPK pathway in A549 cells. The P38 inhibitor SB and the JNK inhibitor SP were applied to A549 cells. After pretreatment with 20 mmol/L of inhibitors, the elevated ATF-2 (downstream of JNK and p38) (A-B) and c-Jun (downstream of JNK) (C-D) activity induced by MSC-CM was significantly decreased. #Indicate significant difference (P < 0.05) versus the LPS group. ## Indicate significant difference (P < 0.01) versus the LPS group. ** Indicate significant difference (P < 0.01) versus the LPS + MSC-CM group.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

7. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 7. Blockage of the JNK and P38 MAPK pathway in primary human small airway epithelial cells (SAEC). The P38 inhibitor SB and the JNK inhibitor SP were applied to SAEC. After pretreatment with 20 mmol/L of inhibitors, the elevated ATF-2 (downstream of JNK and p38) (A-B) and c-Jun (downstream of JNK) (C-D) activity induced by MSC-CM was significantly decreased. #Indicate significant difference (P < 0.05) versus the LPS group. ## Indicate significant difference (P < 0.01) versus the LPS group. ** Indicate significant difference (P< 0.01) versus the LPS + MSC-CM group.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

8. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 8. Blockage of the JNK and P38 MAPK pathway inhibited proliferation and migration of A549 cells stimulated by MSC-CM. A549 cells proliferation under 6µL/mL of MSC-CM with or without pretreatment of 20 mmol/L of SB or SP The MSC-CM induced proliferation (A) and migration (B-E) are partly abrogated by addition of the MAPK inhibitors. ## Indicate significant difference (P < 0.01) versus the LPS group. ** Indicate significant difference (P < 0.01) versus the LPS + MSC-CM group.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0

9. Mesenchymal Stem Cell Conditioned Medium Promotes Proliferation and Migration of Alveolar Epithelial Cells under Septic Conditions <b><i>In Vitro </i></b>via the JNK-P38 Signaling Pathway
Cell Physiol Biochem 2015;37: DOI: /
Fig. 9. Blockage of the JNK and P38 MAPK pathway inhibited proliferation and migration of primary human small airway epithelial cells (SAEC) stimulated by MSC-CM. SAEC proliferation under 6µL/mL of MSC-CM with or without pretreatment of 20 mmol/L of SB or SP The MSC-CM induced proliferation (A) and migration (B, C) are partly abrogated by addition of the MAPK inhibitors. ## Indicate significant difference (P < 0.01) versus the LPS group. ** Indicate significant difference (P < 0.01) versus the LPS+MSC-CM group.
© 2015 S. Karger AG, Basel - CC BY-NC-ND 4.0