The invention relates to a cell sheet construct for neurovascular reconstruction. The cell sheet construct has a vascular endothelial cell layer and a neural stem cell layer, and the two layers are physically in direct contact with each other, where the vascular endothelial cell layer forms branching vasculatures, and the neural stem cell layer differentiates into neurons. The invention also relates to a method for manufacturing the cell sheet construct, having the following steps: culturing vascular endothelial cells on a substrate to form a vascular endothelial cell layer, seeding neural stem cells on the vascular endothelial cell layer to make the neural stem cells be physically in direct contact with the vascular endothelial cell layer, and culturing the neural stem cells and the vascular endothelial cell layer to differentiate into neurons and branching vasculatures to form a cell sheet construct.

The invention relates to a cell sheet construct for neurovascular reconstruction. The cell sheet construct has a vascular endothelial cell layer and a neural stem cell layer, and the two layers are physically in direct contact with each other, where the vascular endothelial cell layer forms branching vasculatures, and the neural stem cell layer differentiates into neurons. The invention also relates to a method for manufacturing the cell sheet construct, having the following steps: culturing vascular endothelial cells on a substrate to form a vascular endothelial cell layer, seeding neural stem cells on the vascular endothelial cell layer to make the neural stem cells be physically in direct contact with the vascular endothelial cell layer, and culturing the neural stem cells and the vascular endothelial cell layer to differentiate into neurons and branching vasculatures to form a cell sheet construct.

Provided are live, artificial, skin substitute products and methods of making and using the same, such as for wound treatment and compound testing, including compound testing for efficacy, toxicity, penetration, irritation and/or metabolism testing of drug candidates or compositions such as cosmetics. Described herein is an artificial mammalian skin substitute product, comprising: (a) optionally, but in some embodiments preferably, a first (“hypodermis-like”) layer comprising live mammalian adipocytes (e.g., induced pre-adipocytes) in a first hydrogel carrier; (b) a second (“dermis-like”) layer contacting or directly contacting the first layer and comprising live mammalian fibroblast cells and' live mammalian follicle dermal papilla cells in combination in a second hydrogel carrier; (c) a third (“epidermis-like”) layer contacting or directly contacting the second layer (i.e., on the opposite side thereof as the first layer, so that the second layer is sandwiched between the first and third layers when the first layer is present), the third layer comprising live mammalian keratinocytes and live mammalian melanocytes in combination in a third hydrogel carrier.

The artificial blood vessel comprises a cortex layer, a fibroblast layer, a smooth muscle cell layer, an endothelial cell layer and an inner cavity. According to the artificial blood vessel, the endothelial layer, the smooth muscle cell layer, the fibroblast layer and the cortex layer are orderly arranged in a three-dimensional space by utilizing integrated technologies of plasma spraying, electrospraying, electrospining, intra-mold pouring and 3D printing; anticoagulant activity of the artificial blood vessel is enhanced by adopting an anticoagulation factor; step-by-step induced differentiation of stem cells in the artificial blood vessel is realized by adopting a growth factor controlled release method; and the artificial blood vessel is cultured by a pulsatile reactor, so that the artificial blood vessel structurally and functionally simulates natural animal blood vessels and provides a corresponding substitute for vascular transplantation and repair.

Engineered human tissue seed constructs are provided that are suitable for implantation in subjects. Methods of making and using the engineered tissue seed constructs are provided.

A method for treating an oral condition of a subject by grafting cultured tissue constructs to the oral tissue. The cultured tissue constructs comprise cultured cells and endogenously produced extracellular matrix components without the requirement of exogenous matrix components or network support or scaffold members. Some tissue constructs of the invention are comprised of multiple cell layers or more than one cell type. The tissue constructs of the invention have morphological features and functions similar to tissues their cells are derived and their strength makes them easily handleable. Preferred cultured tissue constructs of the invention comprise cells derived from human tissue.

An artificial cornea is fabricated by separately culturing live stromal cells, live corneal endothelial cells (CECs) and live corneal epithelial cells (CEpCs), and 3D bioprinting separate stromal, CEC and CEpC layers to encapsulate the cells into separate hydrogel nanomeshes. The CEC layer is attached to a first side of the stromal layer and the CEpC layer to a second side of the stromal layer to define the artificial cornea.

The present invention recognizes that there exists a long felt need for reliable hair growth methods and compositions that do not suffer from side effects and limitations of current technologies, such as surgery using a subject's own hair and pharmaceutical compositions. A first aspect of the present invention is a method of making at least one three dimensional collection of cells capable of forming a functional hair follicle. A second aspect of the present invention is a product produced by the method of making at least one three dimensional collection of cells capable of forming a functional hair follicle of the present invention. A third aspect of the present invention is a method of making at least one functional hair follicle. A fourth aspect of the present invention is a product produced by the method of making at least one functional hair follicle of the present invention. A fifth aspect of the present invention is a method of hair growth in a subject using at least one three dimensional collection of cells capable of forming a functional hair follicle of the present invention. A sixth aspect of the present invention is a method of hair growth in a subject using at least one functional hair follicle of the present invention.

Methods and materials for making complex, living, vascularized tissues for organ and tissue replacement, especially complex and/or thick, structures, such as liver tissue is provided. Tissue lamina is made in a system comprising an apparatus having (a) a first mold or polymer scaffold, a semi-permeable membrane, and a second mold or polymer scaffold, wherein the semi-permeable membrane is disposed between the first and second molds or polymer scaffolds, wherein the first and second molds or polymer scaffolds have means defining microchannels positioned toward the semi-permeable membrane, wherein the first and second molds or polymer scaffolds are fastened together; and (b) animal cells. Methods for producing complex, three-dimensional tissues or organs from tissue lamina are also provided.

Methods of using human airway stem cells in lung epithelial engineering, optionally wherein the cells are contacted with a gamma secretase inhibitor, bioartificial airway organs produced thereby, and the use thereof, e.g., for transplantation. Also methods of treating a bio-artificial matrix with Tenascin-C and/or fibrillin 2.