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 TGF1 (and/or the TGFsignaling pathway) and BMP4 (and/or the BMP signaling pathway).

An optimized method based on a dual promoter vector of the reprogramming factors combined with knock-down of the neural silencing complex RESTi to convert adult fibroblasts into induced neurons (iNs). We have also designed and cloned vector constructs of which some include all these components which allows for a one-step method to efficiently reprogram dermal fibroblasts including those obtained from elderly individuals. The single vector system can be used to obtain iNs of high yield and purity from biopsies from aged individuals with a range of familial and sporadic neurodegenerative disorders including Parkinson's, Huntington's as well as Alzheimer's disease.

Provided is an epithelial tissue stem cell derived from an adult, which lacks at least one tumor suppressor gene.

The present disclosure relates to methods of differentiating pluripotent stem cells or lung progenitor cells into epithelial or basal cells. The present disclosure also relates to epithelial or basal cells made by such methods, organoids containing such epithelial or basal cells, and methods of using the same. The present disclosure also relates to differentiation media for use in the same.

The present disclosure relates to methods of differentiating pluripotent stem cells or lung progenitor cells into alveolar type 2 (AT2) cells. The present disclosure also relates to AT2 cells made by such methods, organoids containing such AT2 cells and methods of using the same. The present disclosure also relates to differentiation media for use in the same.

Described herein are methods and compositions relating to the provision and/or differentiation of certain fully differentiated lung cell types, including surfactant producing cells (e.g. club secretory cells), mucus producing (Goblet) cells, mucus-producing goblet like cells, and beating ciliated cells. In some embodiments, one or more of the differentiated lung cell types is present in an organoid. Differentiation of these lung cell types can be induced by contacting lung stem and/or progenitor cells with one or more inhibitors of H3K9me1/2 methyltransferase, e.g., an inhibitor of G9a and/or Glp (EMHT2 and/or EHMT1).

Described herein are methods and compositions related to induced alveolar epithelial type 2 cells (iAEC2s), e.g., artificially-produced epithelial type 2 cells.

The invention disclosed herein generally relates to methods and systems for growing, expanding and/or obtaining 3-dimensional lung-like epithelium comprising cells having SOX9 protein activity and SOX2+ protein activity. In particular, the invention disclosed herein relates to methods and systems for growing human cells having SOX9 protein activity and SOX2+ protein activity in vitro, and for promoting pluripotent stem cell derived ventral-anterior foregut spheroid tissue into 3-dimensional lung-like epithelium comprising cells having SOX9 protein activity and SOX2+ protein activity.

Various aspects described herein provide methods of generating alveolar or alveolar/airway organoids from a population of lung cells to differentiate into alveolar or alveolar/airway organoids. Also provided herein are methods and compositions for treating lung disease comprising transplantation of the alveolar or alveolar/airway organoids, or a cell isolated therefrom to a subject.

Described herein are methods and compositions relating to the provision and/or differentiation of certain fully differentiated lung cell types, including surfactant producing cells (e.g. club secretory cells), mucus producing (Goblet) cells, mucus-producing goblet like cells, and beating ciliated cells. In some embodiments, one or more of the differentiated lung cell types is present in an organoid. Differentiation of these lung cell types can be induced by contacting lung stem and/or progenitor cells with one or more inhibitors of H3K9me1/2 methyltransferase, e.g., an inhibitor of G9a and/or G1p (EMHT2 and/or EHMT1).