TY - JOUR
T1 - Epithelial to mesenchymal transition (EMT) induced by bleomycin or TFG b1/EGF in murine induced pluripotent stem cell-derived alveolar Type II-like cells
AU - Alipio, Zaida A.
AU - Jones, Nathan
AU - Liao, Wenbin
AU - Yang, Jianchang
AU - Kulkarni, Shilpa
AU - Sree Kumar, K.
AU - Hauer-Jensen, Martin
AU - Ward, David C.
AU - Ma, Yupo
AU - Fink, Louis M.
N1 - Funding Information:
Grants: this project is supported in part by DOD Grants W81XWH-09-1-0451 and W81XWH-10-1-0046 .
PY - 2011/9
Y1 - 2011/9
N2 - Induced pluripotent stem (iPS) cells are derived from reprogrammed somatic cells and are similar to embryonic stem (ES) cells in morphology, gene/protein expression, and pluripotency. In this study, we explored the potential of iPS cells to differentiate into alveolar Type II (ATII)-like epithelial cells. Analysis using quantitative real time polymerase chain reaction and immunofluorescence staining showed that pulmonary surfactant proteins commonly expressed by ATII cells such as surfactant protein A (SPA), surfactant protein B (SPB), and surfactant protein C (SPC) were upregulated in the differentiated cells. Microphilopodia characteristics and lamellar bodies were observed by transmission electron microscopy and lipid deposits were verified by Nile Red and Periodic Acid Schiff staining. C3 complement protein, a specific feature of ATII cells, was present at high levels in culture supernatants demonstrating functionality of these cells in culture. These data show that the differentiated cells generated from iPS cells using a culture method developed previously (Rippon et al., 2006) are ATII-like cells.To further characterize these ATII-like cells, we tested whether they could undergo epithelial to mesenchymal transition (EMT) by exposure to drugs that induce lung fibrosis in mice, such as bleomycin, and the combination of transforming growth factor beta1 (TGF b1) and epidermal growth factor (EGF). When the ATII-like cells were exposed to either bleomycin or a TGF b1-EGF cocktail, they underwent phenotypic changes including acquisition of a mesenchymal/fibroblastic morphology, upregulation of mesenchymal markers (Col1, Vim, a-Sma, and S100A4), and downregulation of surfactant proteins and E-cadherin.We have shown that ATII-like cells can be derived from skin fibroblasts and that they respond to fibrotic stimuli. These cells provide a valuable tool for screening of agents that can potentially ameliorate or prevent diseases involving lung fibrosis.
AB - Induced pluripotent stem (iPS) cells are derived from reprogrammed somatic cells and are similar to embryonic stem (ES) cells in morphology, gene/protein expression, and pluripotency. In this study, we explored the potential of iPS cells to differentiate into alveolar Type II (ATII)-like epithelial cells. Analysis using quantitative real time polymerase chain reaction and immunofluorescence staining showed that pulmonary surfactant proteins commonly expressed by ATII cells such as surfactant protein A (SPA), surfactant protein B (SPB), and surfactant protein C (SPC) were upregulated in the differentiated cells. Microphilopodia characteristics and lamellar bodies were observed by transmission electron microscopy and lipid deposits were verified by Nile Red and Periodic Acid Schiff staining. C3 complement protein, a specific feature of ATII cells, was present at high levels in culture supernatants demonstrating functionality of these cells in culture. These data show that the differentiated cells generated from iPS cells using a culture method developed previously (Rippon et al., 2006) are ATII-like cells.To further characterize these ATII-like cells, we tested whether they could undergo epithelial to mesenchymal transition (EMT) by exposure to drugs that induce lung fibrosis in mice, such as bleomycin, and the combination of transforming growth factor beta1 (TGF b1) and epidermal growth factor (EGF). When the ATII-like cells were exposed to either bleomycin or a TGF b1-EGF cocktail, they underwent phenotypic changes including acquisition of a mesenchymal/fibroblastic morphology, upregulation of mesenchymal markers (Col1, Vim, a-Sma, and S100A4), and downregulation of surfactant proteins and E-cadherin.We have shown that ATII-like cells can be derived from skin fibroblasts and that they respond to fibrotic stimuli. These cells provide a valuable tool for screening of agents that can potentially ameliorate or prevent diseases involving lung fibrosis.
KW - Alveolar Type II cells
KW - Bleomycin
KW - EMT
KW - Epithelial to mesenchymal transition
KW - IPS cells
KW - TGF -EGF
UR - http://www.scopus.com/inward/record.url?scp=80051564941&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=80051564941&partnerID=8YFLogxK
U2 - 10.1016/j.diff.2011.05.001
DO - 10.1016/j.diff.2011.05.001
M3 - Article
C2 - 21596473
AN - SCOPUS:80051564941
SN - 0301-4681
VL - 82
SP - 89
EP - 98
JO - Differentiation
JF - Differentiation
IS - 2
ER -
