Efficient derivation of alveolar type II cells from embryonic stem cells for in vivo application.
In the present study, mouse embryonic stem cells (ESCs) were differentiated into alveolar epithelial type II (AEII) cells for endotracheal injection. These enriched lung-like populations expressed lung epithelial markers SP-A, SP-B, SP-C, and CC10. First we show that rapid differentiation of ESCs requires a dissociated seeding method instead of an embryoid body culture method. We then investigated a two-step differentiation of ESCs into definitive endoderm by activin or A549-conditioned medium as a precursor to lung epithelial cells. When conditioned medium from A549 cells was used to derive endoderm, yield was increased above that of activin alone. Further studies showed that Wnt3a may be one of the secreted factors produced by A549 cells and promotes definitive endoderm differentiation, in part, through suppression of primitive endoderm. Activin and Wnt3a together at appropriate doses with dissociated cell seeding promoted greater endoderm yield than activin alone. Next, fibroblast growth factor 2 was shown to induce a dose-dependent expression of SPC, and these cells contained lamellar bodies characteristic of mature AEII cells from ESC-derived endoderm. Finally, ES-derived lung cells were endotracheally injected into preterm mice with evidence of AEII distribution within the lung parenchyma. This study concludes that a recapitulation of development may enhance derivation of an enriched population of lung-like cells for use in cell-based therapy.
Roszell
Blair
B
School of Biomedical Engineering, Sciences, and Health Systems, Drexel University, Philadelphia, Pennsylvania 19104, USA.
Mondrinos
Mark J
MJ
Seaton
Ariel
A
Simons
Donald M
DM
Koutzaki
Sirma H
SH
Fong
Guo-Hua
GH
Lelkes
Peter I
PI
Finck
Christine M
CM
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology*; D051379:Mice; D011650:Pulmonary Alveoli / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*