Compositions and methods for obtaining functional basal-like cells
The invention disclosed herein generally relates to methods and systems for converting stem cells into specific tissue(s) or cells through directed differentiation. In particular, the invention disclosed herein relates to methods and systems for promoting functional basal-like cells from pluripotent stem cell-derived lung bud tip progenitor organoid tissue through activation of SMAD signaling via activation of TGFβ1 (and/or the TGFβ signaling pathway) and BMP4 (and/or the BMP signaling pathway).
1. A method, comprising:
(i) differentiating human fetal lung bud tip progenitor organoids comprising Sox2 + /Sox9 + cells into organoids comprising cells expressing TP63, wherein the differentiating comprises contacting the human fetal lung bud tip progenitor organoids comprising Sox2 + /Sox9 + cells in vitro with a composition comprising TGF-β and BMP4 to obtain organoids comprising cells expressing TP63; and
(ii) expanding the organoids comprising cells expressing TP63, wherein the expanding comprises contacting the organoids comprising cells expressing TP63 with a composition comprising FGF10, Y27632, A8301, and Noggin;
wherein the cells expressing TP63 further express one or more of KRT5, KRT14, and F3;
wherein the expanded organoids comprising cells expressing TP63 are capable of clonal expansion, self-renewal and multilineage differentiation.