Abstract: A method for producing a multi-tissue organoid generally includes obtaining pluripotent stem cells (PSCs) and inducing growth of the multi-tissue organoid by culturing the harvested PSCs in suspension in a culture vessel. Another method for producing a multi-tissue organoid generally includes introducing a pluripotent stem cell into a cell culture medium comprising hyaluronic acid, transferring the pluripotent stem cell to a cell culture device that does not include a three-dimensional matrix, culturing the pluripotent stem cell in the cell culture device for at least 1 week, and producing a multi-tissue organoid comprising cartilage, bone, fibrous connective tissue, brain tissue, or epithelial tissue, or a combination thereof.

Abstract: Disclosed herein are methods of producing a three-dimensional cell culture microenvironment, which may include exposing a lyophilized polycation coupon to a polyanion and a fluid medium. The lyophilized polycation coupon may optionally include a void volume, and the polyanion may optionally be contained within a slurry. Such methods may also include the steps of evaporating off the fluid medium of the slurry to produce a dried three-dimensional architecture, administering a hydrating fluid to the dried three-dimensional architecture, and producing the three-dimensional cell culture microenvironment. Also disclosed herein are three-dimensional cell culture microenvironments formed by various processes, as well as kits which include such three-dimensional cell culture microenvironments provided in a multi-well plate, and a hydrating fluid.