A composition including, (a) a mineral particle, (b) endothelial cells and mesenchymal cells, and (3) hyaluronic acid, is provided. Moreover, a kit which includes: a syringe, a mineral particle covered with endothelial cells and mesenchymal cells organized in 2 or more cell layers attached to the mineral particle, and hyaluronic acid, is also provided. Last, a method for filling a gap in a bone of a subject in need thereof, including contacting the gap with a composition of: (a) a mineral particle, (b) endothelial cells and mesenchymal cells, and (3) hyaluronic acid is provided.

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.

Methods are disclosed for fabricating a three-dimensional engineered blood retinal barrier (BRB) comprising a choroid and retinal pigment epithelial cells. The methods include the use of bioprinting. Also disclosed is a three-dimensional engineered BRB, and its use. Methods are also disclosed for using the three-dimensional engineered BRB, such as for the treatment of retinal degeneration in a subject or screening. A three-dimensional printing insert that is adapted for bioprinting on a culture substrate sheet that is securely retained within and exposed through a printing frame is also disclosed.

A method for preparing zonal layered chondrocyte sheets, comprising the steps: (a) providing a cartilage sample from a subject; (b) isolating chondrocytes from the cartilage sample and then isolating superficial zone chondrocytes, middle zone chondrocytes and deep zone chondrocytes from the chondrocytes; (c) culturing the deep zone chondrocytes until reaching 100% cell confluence to form a deep zone chondrocyte sheet; (d) seeding the middle zone chondrocytes on the top of the cultured deep zone chondrocyte sheet from the step (c) and culturing the middle zone chondrocytes until reaching 100% cell confluence to form a middle zone chondrocyte sheet; and (e) seeding the superficial zone chondrocytes on the top of the cultured middle zone chondrocyte sheet from the step (d) and culturing the superficial zone chondrocytes until reaching 100% cell confluence to form a superficial zone chondrocyte sheet to obtain the zonal layered chondrocyte sheets.

The present invention provides, in some embodiments, multi-layer silk compositions including a first layer comprising silk fibroin and keratinocytes, a second layer comprising silk fibroin and fibroblasts, a third layer comprising silk fibroin and adipocytes, and a plurality of nervous system cells, wherein at least some of the plurality of nervous system cells span at least two layers, and methods of making and using the same. In some embodiments, provided methods and compositions further include immune cells and/or endothelial cells.

The present specification relates to a method for manufacturing a multilayered cell sheet, and a multilayered cell sheet manufactured using same, the method comprising the steps of: (a) forming a first cell layer on a first substrate, which has a melting point or is changed from being hydrophobic to being hydrophilic at any one temperature from 0° C. to 30° C.; (b) forming a second cell layer on a second substrate to be degraded by an enzyme; (c) making the first cell layer and the second cell layer come in contact with each other; (d) selectively removing the first substrate by providing a temperature lower than or equal to the melting point of the first substrate or a temperature at which the first substrate is changed to being hydrophilic; and (e) selectively removing the second substrate by making the second substrate come in contact with a solution containing the enzyme.

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.

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.

The present invention provides a method of producing a cell sheet comprising a retinal pigment epithelial cell layer and a vascular forming cell layer composed of stem cells having vascular formation ability or blood vessel constituting cells, comprising a step of laminating the retinal pigment epithelial cell layer and the vascular forming cell layer, and a cell sheet obtained by said method. Furthermore, the present invention provides a cell sheet for transplantation, comprising a cell layer formed with retinal pigment epithelial cells obtained by inducing differentiation of stem cells or progenitor cells ex vivo, a basement membrane secreted from said cells, and a vascular forming cell layer composed of stem cells having vascular formation ability or blood vessel constituting cells.

The invention provides a three dimensional (3D), porous, biodegradable and biocompatible tissue engineering scaffold, wherein at least 25% w/w of the scaffold is particulate egg shell membrane (ESM) distributed substantially uniformly therein and the scaffold is essentially dry. Methods for preparing the same by freeze-drying and cryogelation and the use thereof in methods of tissue engineering and to promote the healing of wounds are also provided.