Provided herein are floating hydrogel droplet culture methods that enable scaling of stem cell derived alveolar epithelial cell (AEC) expansion to numbers compatible with large animal or human whole lung engineering, as well as molds for generating the droplets and methods of use thereof.

Disclosed herein is a method for cultivating primary human pulmonary alveolar epithelial cells (HPAEpiC), which includes cultivating the primary HPAEpiC in a first medium containing a basal medium, a culture supplement, and a Rho kinase inhibitor, and a second medium containing the basal medium and the culture supplement in sequence. The culture supplement includes Jagged-1 (JAG-1) peptide, human Noggin protein, transforming growth factor-β (TGF-β) type I receptor inhibitor SB431542, human fibroblast growth factor 7 (hFGF-7), hFGF-10, and glycogen synthase kinase 3 (GSK-3) inhibitor CHIR99021. Also disclosed is a method for preparing a three-dimensional cell culture of alveolar epithelium using the first medium and the second medium.

Compositions useful for propagation of pluripotent stem cells are provided. The compositions comprise a polysaccharide hydrogel linked to a peptide fragment of the extracellular domain of epithelial cadherin. Methods of making the composition, and culturing pluripotent stem cells also are provided.

The present disclosure relates to a 3D bioprinter (1) comprising a base unit (2). The base unit (2) has a support (3) adapted for mounting of at least one toolhead (4), a communication interface part (5) for communication of data with the at least one toolhead (4), when mounted, and a base unit processing element (7) adapted to communicate with a toolhead processing element (8) of the at least one toolhead over said communication interface part (5). The present disclosure relates further to a 3D bioprinter toolhead. The present disclosure relates further to a method for bioprinting a construct.

Sub-millimeter scale three-dimensional (3D) structures are disclosed with customizable chemical properties and/or functionality. The 3D structures are referred to as drop-carrier particles. The drop-carrier particles allow the selective association of one solution (i.e., a dispersed phased) with an interior portion of each of the drop-carrier particles, while a second non-miscible solution (i.e., a continuous phase) associates with an exterior portion of each of the drop-carrier particles due to the specific chemical and/or physical properties of the interior and exterior regions of the drop-carrier particles. The combined drop-carrier particle with the dispersed phase contained therein is referred to as a particle-drop. The selective association results in compartmentalization of the dispersed phase solution into sub-microliter-sized volumes contained in the drop-carrier particles. The compartmentalized volumes can be used for single-molecule assays as well as single-cell, and other single-entity assays.

The present disclosure provides a method of fabricating curvature-defined (C-D) or shape-defined (S-D) concave and convex polydimethylsiloxane (PDMS) surfaces and a method of fabricating C-D or S-D convex and concave gel surfaces for use in cell and tissue culturing and in other surface and interface applications, and provides a method of using C-D or S-D convex and concave surfaces with varying curvatures to direct cell attachment, spreading, and migration.

The present invention relates to a method of expanding pluripotent stem cells (PSC) in suspension culture in a bioreactor, the method comprising (i) adding an inhibitor of ROCK (ROCKi) to pluripotent stem cells being cultivated in suspension in the bioreactor; (ii) adding a cell dissociation agent, thereby dissociating aggregates of the pluripotent stem cells; (iii) diluting the cell dissociation agent added in step (ii) by adding an excess volume of culture medium sufficient to decrease the concentration of the cell dissociation agent to a concentration at which cell aggregates can form again; and (iv) culturing of the mixture obtained in step (iii) under suitable conditions that allow the expansion of the PSCs.

A bioartificial device, such as a bioartificial pancreas, for implantation in a patient's vascular system. The bioartificial pancreas includes a scaffold adapted to engage an interior wall of a blood vessel, a cellular complex support by the scaffold and extending longitudinally within the interior cavity of the scaffold so as to be exposed to the blood flow when the scaffold is engaged with the blood vessel, the cellular complex support comprising one or more pockets bordered by thin film; and cellular complex comprising pancreatic islets disposed in the one or more pockets, the thin film being adapted to permit oxygen and glucose to diffuse from flowing blood into the one or more pockets at a rate sufficient to support the viability of the islets. The invention also includes methods of making and using a bioartificial pancreas.

The present invention provides a scaffold for culturing retinal tissue comprising an amount of gelatin, an amount of chondroitin sulfate, an amount of hyaluronic acid, wherein the amount of gelatin, chondroitin sulfate, and hyaluronic acid are prepared into a three-dimensional monolith, wherein the monolith is sectioned into planar sheets, and an amount of laminin-521.

The invention provides a construct (1) comprising a number N of material types (100, 110, . . . ), wherein N is at least 2, wherein at least two of the material types (100, 110, . . . ) comprise granular material (101) comprising particles (10), wherein the granular material (101) at least defines an exterior surface (6) of the construct (1), wherein the construct (1) is self-supporting, and wherein the construct (1) is (i) self-healing or is (ii) configured for being self-healing by changing a liquid (15) content of the construct (1); wherein the different material types (100, 110, . . . ) mutually differ in at least one characteristic (19) selected from the group consisting of a physical characteristic and a chemical characteristic.