Provided is a method for making a biotransplant, comprising introducing autologous fibroblasts isolated from an oral mucosa of a patient into a platelet-rich fibrin (PRF) membrane using linear retrograde needle injection, where a needle is inserted into a thickness of the PRF membrane and a first puncture is made and then a series of punctures are made that are linearly aligned or arrayed with the first puncture and are spaced at a predetermined distance from a prior puncture. A method of treatment of a periodontal tissue and a biotransplant comprising a PRF membrane and autologous fibroblasts are also provided.

The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated/progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and/or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth/regeneration.

A system and method for printing cells in a medium. A multi-dimensional printer, stably constructed of low-mass parts, can include a computer numerically controlled system that can enable motors driving delivery systems. The motors can include encoders that can enable achieving arbitrary resolution. The motors can drive ballscrews to enable linear motion of delivery systems, and the delivery systems can enable printing of a biological material in a pre-selected pattern in a petri dish. The petri dish can accommodate a medium such as a gel, and can further accommodate a vision system that can detect actual position and deflection of the delivery system needle. The printer can accommodate multiple delivery systems and therefore multiple needles of various sizes.

The present disclosure provides, in various aspects, a method of forming a prefabricated innervated pre-vascularized pre-laminated (PIPP) flap having a stoma or lumen. The method includes providing a cell construct including skin cells and/or mucosa cells. The method further includes forming an integrated in vivo composite at a donor site by grafting the cell construct onto a muscle. The method further includes stabilizing the composite on an obturator component. The method further includes developing a microvascular system in the composite by retaining it in vivo at the donor site for a predetermined period of time. The method further includes removing the obturator component from the stoma or lumen. In certain aspects, the present disclosure also provides a method of restoring a defect including damaged or surgically removed soft tissue using a PIPP flap. In certain aspect, the present disclosure also provides an obturator component for maintaining the stoma or lumen.

The subject invention is directed to a mixed cell composition to generate a therapeutic protein at a target site by providing a first population of mammalian cells transfected or transduced with a gene that is sought to be expressed, and a second population of mammalian cells that have not been transfected or transduced with the gene, wherein endogenously existing forms of the second population of mammalian cells are decreased at the target site, and wherein generation of the therapeutic protein by the first population of mammalian cells at the target site stimulates the second population cells to induce a therapeutic effect.

Provided herein are constructs of micro-aggregate multicellular, minimally polarized grafts containing Leucine-rich repeat-containing G-protein coupled Receptor (LGR) expressing cells for wound therapy applications, tissue engineering, cell therapy applications, regenerative medicine applications, medical/therapeutic applications, tissue healing applications, immune therapy applications, and tissue transplant therapy applications which preferably are associated with a delivery vector/substrate/support/scaffold for direct application.

Compositions and methods of transplanting cells by grafting strategies into solid organs (especially internal organs) are provided. These methods and compositions can be used to repair diseased organs or to establish models of disease states in experimental hosts. The method involves attachment onto the surface of a tissue or organ, a patch graft, a “bandaid-like” covering, containing epithelial cells with supporting early lineage stage mesenchymal cells. The cells are incorporated into soft gel-forming biomaterials prepared under serum-free, defined conditions comprised of nutrients, lipids, vitamins, and regulatory signals that collectively support stemness of the donor cells. The graft is covered with a biodegradable, biocompatible, bioresorbable backing used to affix the graft to the target site. The cells in the graft migrate into and throughout the tissue such that within a couple of weeks they are uniformly dispersed within the recipient (host) tissue. The mechanisms by which engraftment and integration of donor cells into the organ or tissue involve multiple membrane-associated and secreted forms of MMPs.

Compositions and methods are provided for lamellar and defect reconstruction of corneal stromal tissue using biomaterials that form a defined hydrogel structure in situ, including interpenetrating (IPN) and semi-IPN hydrogels.

A method may comprises bringing cells suspended in an aqueous medium into contact with a plurality of fragmented collagen pieces and, after the cells brought into contact with the plurality of fragmented collagen pieces and the plurality of fragmented collagen pieces are concentrated, culturing the cells brought into contact with the fragmented collagen pieces, with the plurality of fragmented collagen pieces, to form a three-dimensional tissue.

A method for decellularization of a skin tissue according to an embodiment of the present invention comprises: a step of preparing a skin tissue to be decellularized; a peeling preparation step of treating the skin tissue with a first solution containing trypsin; and a peeling step of removing subcutaneous fat from the skin tissue after the peeling preparation step.