Efficient derivation of purified lung and thyroid progenitors from embryonic stem cells

Abstract

During early lung and thyroid development two populations of Nkx2-1-expressing progenitors in the foregut endoderm give rise to the entire lung and thyroid epithelium. To date, little is known about the genetic programs of these progenitors since they are few in number, present only fleetingly in the embryo, and have not been studied in pure preparations. We demonstrate here the purification, and directed differentiation in culture of primordial lung and thyroid progenitors derived from mouse embryonic stem (ES) cells. We find that sequential stage-specific and time-dependent inhibition of Transforming Growth Factor β and Bone Morphogenetic Protein signaling, followed by combinatorial stimulation of BMP and FGF signaling is necessary for the efficient specification of these cells from definitive endodermal precursors in vitro. Employing a novel Nkx2-1GFP knock-in reporter ES cell line, these progenitors can be isolated, expanded in culture, and display a global transcriptome that overlaps that of an in vivo developing lung epithelium. Moreover, these progenitors can be induced to sequentially express a broad repertoire of lineage markers indicative of the full diversity of lung and thyroid epithelial differentiation. Thus, we have derived the first population of progenitors that recapitulates the developmental milestones of lung/thyroid development.