Compositions and methods for producing major ocular cell types, including retinal ganglion cells, photoreceptors, retinal pigmented epithelium and corneal endothelial cells, from human pluripotent stem cells under defined culture conditions are provided.

The present invention contemplates compositions, devices and methods of simulating biological fluids in a fluidic device, including but not limited to a microfluidic chip. In one embodiment, fluid comprising a colloid under flow in a microfluidic chip has a fluid density or viscosity similar to a bodily fluid, e.g. blood, lymph, lung fluid, or the like. In one embodiment, a fluid is provided as a Theologically biomimetic blood surrogate or substitute for simulating physiological shear stress and cell dynamics in fluidic device, including but not limited to immune cells.

Ischemic conditions are a leading cause of death for both men and women. Ischemia, a condition characterized by reduced blood flow and oxygen to an organ. Re-establishment of blood flow, or reperfusion, and re-oxygenation of the affected area following an ischemic episode is critical to limit irreversible damage. However, reperfusion also associates potentially damaging consequences. For instance, increased vascular permeability is an important contributor to edema and tissue damage following ischemic events. Here the inventors shows that genetic inhibition of PI3K-C2β reduces cerebral infarction in two ischemia/reperfusion (I/R) models and improves neurological outcome. The genetic inhibition stabilizes the blood—brain barrier (BBB) after ischemic stroke and reduces inflammation. Accordingly, the present invention relates to a method for the preservation of vascular endothelial cell barrier integrity in a patient in need thereof comprising administering to the subject a therapeutically effective amount of a PI3KC2β inhibitor.

An in vitro microfluidic intestine on-chip is described herein that mimics the structure and at least one function of specific areas of the gastrointestinal system in vivo. In particular, a multicellular, layered, microfluidic intestinal cell culture, which is some embodiments is derived from patient's enteroids-derived cells, is described comprising L cells, allowing for interactions between L cells and gastrointestinal epithelial cells, endothelial cells and immune cells. This in vitro microfluidic system can be used for modeling inflammatory gastrointestinal autoimmune tissue, e.g., diabetes, obesity, intestinal insufficiency and other inflammatory gastrointestinal disorders. These multicellular-layered microfluidic intestine on-chips further allow for comparisons between types of gastrointestinal tissues, e.g., small intestinal duodenum, small intestinal jejunum, small intestinal ileum, large intestinal colon, etc., and between disease states of gastrointestinal tissue, i.e. healthy, pre-disease and diseased areas. Additionally, these microfluidic gut-on-chips allow identification of cells and cellular derived factors driving disease states and drug testing for reducing inflammation.

The present disclosure provides a device for patterning extracellular matrix (ECM) hydrogel comprising a first layer surface patterned to define a microchannel, a second layer comprising a loading channel in fluid communication with loading ports to receive an ECM hydrogel, wherein the first layer is attached over the second layer such that the patterned surface faces the loading channel to define an open chamber with regions of reduced cross-sectional area, and wherein the ECM hydrogel is confined to fill said regions, thereby forming a perfusable channel in the open chamber. The present disclosure also provides the same device wherein the second layer is a substrate without a loading channel and is optically pervious; and additionally provides a method of patterning ECM hydrogel comprising use of the aforementioned device. Importantly, ECM patterning is achieved by surface tension between the ECM hydrogel and the first layer at the boundaries of the microchannel.

The present invention relates to the identification of ALMS1 as the missing player involved in the regulation of the insulin-mediated glucose uptake through GLUT4 sorting vesicles, and to the down-regulation of ALMS1 by αPKC. Accordingly, the present invention relates to a molecule capable of preventing the binding of αPKC on ALMS1 for use for treating or preventing diabetes, in particular type 2 diabetes. In addition, the present invention relates to a method for identifying molecule capable of preventing the binding of αPKC on ALMS1.

This document provides materials and methods for making and using functional (e.g., vascularized) organ models (e.g., pancreas models). For example, functional pancreas models including an ECM containing a plurality (e.g., two or more) of pancreatic islets, and a vascular network are provided.

Disclosed is a method for producing an in vitro model for blood-brain barrier, including (a) a culturing conditionally immortalized astrocytes on one surface of a porous membrane and culturing conditionally immortalized brain pericytes on the other surface of the porous membrane, until both of the cells become a sheet; (b) culturing conditionally immortalized brain microvascular endothelial cells in a culture vessel, until the cells become a sheet; (c) peeling off the sheet of conditionally immortalized brain microvascular endothelial cells; (d) allowing the sheet of conditionally immortalized brain microvascular endothelial cells to come into contact with the sheet of conditionally immortalized brain pericytes, so that the sheets are arranged in layers; and (e) co-culturing a cell culture comprising three layers consisting of the sheet of conditionally immortalized brain microvascular endothelial cells, the sheet of conditionally immortalized brain pericytes, and the sheet of conditionally immortalized astrocytes.

Compositions, methods, and kits are provided for determining whether a subject is at risk of developing insulin resistance. In particular, phosphorylated Akt, reactive oxygen species (ROS), SIRT1, eNOS, CDH13, IRS1 and NO production have been identified as biomarkers associated with insulin resistance and type 2 diabetes. The diagnostic methods comprise measuring the level of at least one biomarker in induced pluripotent stem cells derived from somatic cells of the subject, which have been differentiated into endothelial cells (IPSC-ECs).

Methods fabricate an endothelialized myocardium usable for screening of a drug. A microfibrous hydrogel scaffold is manufactured with additive manufacturing that concurrently bioprints endothelial cells directly within the microfibrous hydrogel scaffold. A bioink is bioprinted into an arrangement of one or more microfibers. The bioink includes at least one crosslinking component and suspended endothelial cells. The crosslinking component or components are crosslinked to yield the microfibrous hydrogel scaffold having the endothelial cells embedded directly within. The microfibrous hydrogel scaffold is seeded with cardiomyocytes to yield the endothelialized myocardium with a controlled anisotropy. The endothelialized myocardium can be incubated until the endothelialized myocardium matures into spontaneously beating myocardial tissue having contractions aligned with the controlled anisotropy. The beating myocardial tissue can be used to screen a drug when the beating myocardial tissue is embedded within a microfluidic perfusion bioreactor.