The present disclosure relates to novel multi-organ-chips establishing the differentiation of induced pluripotent stem cell (iPSC)-derived cells into organ equivalents on microfluidic devices and corresponding methods of generating organ equivalents. The present disclosure also relates to novel bioengineered tissue constructs mimicking organ barriers generated with iPSC-derived endothelial cells and/or organoids bioprinted in, and/or seeded on, a hydrogel. The present disclosure further relates to methods of bio-engineering organ constructs comprising co-culturing iPSC-derived organ precursor cells and iPSC-derived fibroblasts and endothelial cells. The present disclosure specifically provides a microfluidic device comprising: (i) iPSC-derived hepatocyte precursor cells; (ii) iPSC-derived intestinal precursor cells; (iii) iPSC-derived renal tubular precursor cells; and (iv) iPSC-derived neuronal precursor cells; wherein the iPSC-derived precursor cells according to (i), (ii), (iii) and (iv) are differentiated from a single donor iPSC reprogrammed from a single type of somatic cell.

Provided herein according to some embodiments is an in vitro construct useful as a model for a hematopoietic microenvironment, which may include: a microfluidic device having multiple chambers; and two or more populations of cells (e.g., 3 or 4 populations of cells) (or niches) selected from: 1) mesenchymal cells (e.g., Stro-1+; MSC); 2) osteoblasts (OB; optionally said osteoblasts provided by differentiating mesenchymal cells to differentiated osteoblasts); 3) arterial endothelium (e.g., CD146+NG2+; AEC); and 4) sinusoidal endothelium (CD146+NG2; SEC), wherein each of said two or more populations of cells are provided in a separate chamber of the microfluidic device. Methods of making and using the construct are also provided.

Methods for using vibration to harvest cells grown in 3D culture are provided. The methods entail the application of force cells attached to a 3D matrix of sufficient amplitude, frequency, and duration to detach cells from the matrix and to flush the detached cells out of the matrix material. An apparatus for performing the methods of the invention as provided.

The invention relates to a method of culturing an epithelial cell on a solid surface.

The present disclosure provides a system, including methods and apparatus, for culturing, monitoring, and/or analyzing organoids. In an exemplary method of organoid culture, the method may comprise disposing a scaffold in a receptacle having an open side. A sealing member may be bonded to the open side of the receptacle to create a chamber. An organoid may be formed in the chamber using the scaffold. Fluid and/or at least one substance may be introduced into the chamber from an overlying reservoir for contact with the organoid.

An in vitro microfluidic organ-on-chip is described herein that mimics the structure and at least one function of specific areas of the epithelial system in vivo. In particular, a multicellular, layered, microfluidic culture is described, allowing for interactions between lamina propria-derived cells and the associated tissue specific epithelial cells and endothelial cells. This in vitro microfluidic system can be used for modeling inflammatory tissue, e.g., autoimmune disorders involving epithelia and diseases involving epithelial layers. These multicellular, layered microfluidic organ-on-chip, e.g. epithelia-on-chip further allow for comparisons between types of epithelia tissues, e.g., lung (Lung-On-Chip), bronchial (Airway-On-Chip), skin (Skin-On-Chip), cervix (Cervix-On-Chip), blood brain barrier (BBB-On-Chip), etc., in additional to neurovascular tissue, (Brain-On-Chip), and between different disease states of tissue, i.e. healthy, pre-disease and diseased areas. Additionally, these microfluidic organ-on-chips allow identification of cells and cellular derived factors driving disease states in addition to drug testing for reducing inflammation effecting epithelial regions.

Aspects of the present disclosure include methods that include co-culturing a cell and a microparticle that includes a capture ligand, in a culture medium under conditions in which a biomarker produced by the cell is bound by the capture ligand. Such methods may further include detecting (e.g., by flow or mass cytometry) complexes that include the microparticle, the capture ligand, the biomarker, and a detection reagent. The methods may further include determining the proportion or number of cells among a heterogeneous cell population that produced the biomarker and/or the level of biomarker secreted by such cells. Compositions, systems and kits are also provided.

The present invention provides hollow microcarriers for cell culture. The hollow microcarriers form a shell around a hollow interior and can be opened to permit cell infiltration or harvesting. The hollow microcarriers protect cells from hydrodynamic shear stress without hindering the diffusion of nutrients in and out of their hollow interior.

A cell culture article includes a substrate having a polymer coating that is conducive to colony passaging of cells cultured on the coating. Example polymer coatings are formed from polygalacturonic acid (PGA), alginate, or combinations thereof. Cells cultured on the polymer coating can be separated from the substrate as a colony or layer of cells by exposing the polymer coating to (i) a chelating agent, (ii) a proteinase-free enzyme, or (iii) a chelating agent and a proteinase-free enzyme.

Embodiments of the present invention are directed to microscale and millimeter scale tissue scaffolding structures that may be static or expandable and which may be formed of biocompatible metals or other materials that may be coated to become biocompatible. Scaffold structures may include features for holding desired biological or physiological materials to enhance selected tissue growth. Scaffolding devices may be formed by multi-layer, multi-material electrochemical fabrication methods.