Described are DNA molecules which can be transcribed into an mRNA harbouring novel UTR sequences combining the advantages of being extremely short and at the same time allowing for high translation efficiencies of RNA molecules containing them. Further, described are vectors comprising such a DNA molecule and to host cells comprising such a vector. Moreover, described are corresponding RNA molecules containing such UTRs. Further, described in a pharmaceutical composition comprising the described RNA molecule are optionally a pharmaceutically acceptable carrier as well as to the use of the described UTRs for translating a coding region of an RNA molecule into a polypeptide or a protein encoded by said coding region.

Disclosed is an in-vitro protocol for differentiating human induced pluripotent stem cells (hiPSCs) or human embryonic stem cells (hESC) to give rise to a definitive endoderm, followed by progression into anteriorized foregut endoderm that has the ability to give rise to both proximal and distal lung epithelial cells. The protocol not only offers great opportunities for the study of human development but also have tremendous potential for future clinical cell-based therapies. The protocol outlined here is used to differentiate hiPSCs into lung epithelial cell types through a process that faithfully recapitulates the stepwise events observed in-vivo. The was followed with the working cell bank of an hiPSC line made under current Good Manufacturing Practice (cGMP) conditions, a necessary step for the future clinical application of these cells.

The present disclosure provides methods for generating lung progenitor cells, and populations of cells made using the methods. The lung progenitors and related compositions can be used as therapeutic treatments for various pulmonary disorders or related injuries.

In vitro equine organ model systems, and methods of making and using such systems, are provided and can include an organoid prepared using equine tissue associated with the organ of interest; or equine primary cells, wherein the equine primary cells are derived from equine tissue associated with an organ of interest, or derived from an organoid prepared using equine tissue associated with the organ of interest.

Abstract: We describe a robust human adult distal lung organoid method with a procedure for everting organoids to essentially turn them inside out. This then relocates the apical ACE2-expressing surfaces of cells to the organoid exterior, where they can then be easily infected by SARS-CoV-2 added to the tissue culture medium. Further, this method can be used for infection of any distal lung pathogen that infects apically. Alternatively, if a pathogen interacts basolaterally then eversion is not necessary, and the human adult distal lung organoids can be infected as is.

A reagent for differentiating somatic cells into alveolar epithelial cells includes an NK2 homeobox family gene expression vector, and a Fox family gene expression vector, a method for manufacturing alveolar epithelial cells, the method including forcing a somatic cell to express an NK2 homeobox family gene and a Fox family gene and culturing the somatic cell after forced expression, an alveolar epithelial cell manufactured by the manufacturing method, and a cell medicine including the alveolar epithelial cell.

Provided herein are methods and systems for bio-printing of three-dimensional cell-containing matrixes. Further, provided herein are methods and systems for generating a three-dimensional (3D) structure corresponding to a biological material, such as a kidney or lung comprising either nephron or alveolar structures. Also provided herein are bio-printed three-dimensional matrices for use in the generation nephron and/or alveolar structures.

The present invention relates to the discovery that different stem cell types (e.g., bone marrow-derived mesenchymal stem cells (RM-MSC) and adipose-derived mesenchymal stem cells (AT-MSC)) undergo large changes in lung epithelial marker 5 expression depending on the substrate on which they are cultured. The present invention includes methods and compositions for differentiating of mesenchymal stem cells, such as bone marrow and adipose tissue mesenchymal stem cells, into lung cells, populations of lung cells, and methods of alleviating or treating a lung defect in a subject in need thereof.

Provided herein are methods and compositions relating, in part, to the generation of human progenitor cells committed to the lung lineage and uses of such cells for treatment of lung diseases/disorders or injury to the lung. Whether an adult stem cell can be isolated from human adult lung remains controversial in the art and at present, methods for isolating and using adult lung stem cells from humans lack reproducibility. Thus, the methods and compositions described herein are advantageous over the present state of knowledge in the art and permit the generation of human lung progenitor cells for treatment, tissue engineering, and screening assays.

The presently disclosed subject matter provides a microfluidic device that can simulate the cross section of the large and small human airways, including the air-exposed epithelial layer, the adjacent surrounding stromal layer, and the blood-facing endothelial layer of near-by vessels in the circulatory system. The microfluidic device can reconstitute the air-liquid interface in the lung and molecular transport characteristics of bronchi and bronchioles in the human pulmonary airways, and provide a more realistic alternative to current in vitro models of airway structures. Additionally, the model can reconstitute the native response of airway tissues to infection by bacterial and viral agents, and also the extravasation of immune cells from the bloodstream and into the stromal and epithelial compartments of the lung in response to an infection. The presently disclosed subject matter also provides microfluidic devices that include multiple chambers assembled by layered stacking or bonding of a basal chamber, a first membrane, an interstitial chamber, a second membrane and an apical chamber.