2092results about "3D culture" patented technology

A stromal cell-based three-dimensional cell culture system is prepared which can be used to culture a variety of different cells and tissues in vitro for prolonged periods of time. The stromal cells and connective tissue proteins naturally secreted by the stromal cells attach to and substantially envelope a framework composed of a biocompatible non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells. The living stromal tissue so formed provides the support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture and / or cultures implanted in vivo. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts in vivo, which can be utilized in the body as a corrective tissue. For example, and not by way of limitation, the three-dimensional cultures can be used to form tubular tissue structures, like those of the gastrointestinal and genitourinary tracts, as well as blood vessels; tissues for hernia repair and / or tendons and ligaments; etc.

A process and apparatus are provided for manufacturing complex parts and devices which utilize a CAD environment to design a part or device to be created (FIG. 1); Boolean, scaling, smoothing, mirroring, or other operations to modify the CAD design; a software interface to convert the CAD designed part (Data Process System) or device into a heterogeneous material and multi-part assembly model (Design Input Model) which can be used for multi-nozzle printing; and a multi-nozzle system to print the designed part or device using different, specialized nozzles (Tissue substitutes).

A device, method and process for three-dimensional spatial localization and functional interconnection of the same or different types of cells. The two or three-dimensional device comprising multiple layers containing wells for cell deposition where both the wells and layers are interconnected through microfluidic channels. A process for fabricating the three-dimensional device and a method for depositing different types of cells within the device in a functional interdependent spatial orientation thereby mimicking physiological functions. The device is useful for diagnostic assays, determination of dysfunction of certain cells in the system, quantification of production of cellular proteins, metabolites, hormones or other cellular products, for organ or tissue replacement, for co-culturing different cells, for testing pharmaceutical agents and as a bioreactor for production of biologicals.

A 3-dimensional structure comprising suitable cells (or entities) and the ECM (or matrix) that has been completely produced and arranged by these cells (or entities) that promotes the differentiation, dedifferentiation and / or transdifferentiation of cells and / or formation of tissue in vitro and in vivo, while at the same time promoting cell growth, proliferation, migration, acquisition of in vivo-like morphology, or combinations thereof, and that 1. provides structural and / or nutritional support to cells, tissue, organs, or combinations thereof, termed an “Autogenic Living Scaffold” (ALS); or 2. is capable of being transformed into a more complex tissue (or matrix) or a completely different type of tissue (or matrix), termed a “Living Tissue Matrix” (LTM). Autogenic means it is self-produced. The living cells that produce the LTM or ALS, or are added to Autogenic Living Scaffolds, may be genetically engineered or otherwise modified. The matrix component of the ALS or LTM provides a structural framework for cells that guide their direction of growth, enables them to be correctly spaced, prevents overcrowding, enables cells to communicate between each other, transmit subtle biological signals, receive signals from their environment, form bonds and contacts that are required for proper functioning of all cells within a unit such as a tissue, or combinations thereof. The ALS or LTM may thus provide proper or supporting mechanical and chemical environments, signals, or stimuli to other cells, to the cells that produce the ALS, to surrounding tissue at an implantation site, to a wound, for in vitro and ex vivo generation and regeneration of cells, tissue and organs, or combinations thereof. They may also provide other cells with nutrients, growth factors, and / or other necessary or useful components. They may also take in or serve as buffers for certain substances in the environment, and have also some potential at adapting to new environments.

This disclosure provides an improved system for culturing human pluripotent stem cells. Traditionally, pluripotent stem cells are cultured on a layer of feeder cells (such as mouse embryonic fibroblasts) to prevent them from differentiating. In the system described here, the role of feeder cells is replaced by components added to the culture environment that support rapid proliferation without differentiation. Effective features are a suitable support structure for the cells, and an effective medium that can be added fresh to the culture without being preconditioned by another cell type. Culturing human embryonic stem cells in fresh medium according to this invention causes the cells to expand surprisingly rapidly, while retaining the ability to differentiate into cells representing all three embryonic germ layers. This new culture system allows for bulk proliferation of pPS cells for commercial production of important products for use in drug screening and human therapy.

A stromal cell-based three-dimensional cell culture system is provided which can be used to culture a variety of different cells and tissues in vitro for prolonged periods of time. The stromal cells along with connective tissue proteins naturally secreted by the stromal cells attach to and substantially envelope a framework composed of a biocompatible non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells. Living stromal tissue so formed provides support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture and / or cultures implanted in vivo. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts in vivo, which can be utilized in the body as a corrective tissue. The three-dimensional cultures can be used to form tubular tissue structures, like those of the gastrointestinal and genitourinary tracts, as well as blood vessels; tissues for hernia repair and / or tendons and ligaments. A three-dimensional filamentous tissue having tendon or ligament function is prepared containing fibroblasts and collagen naturally secreted by the fibroblasts attached to and substantially enveloping a three-dimensional filamentous framework.

This invention provides a fibrin sealant bandage, wherein said fibrin sealant may be supplemented with at least one composition selected from, for example, one or more regulatory compounds, antibody, antimicrobial compositions, analgesics, anticoagulants, antiproliferatives, anti-inflammatory compounds, cytokines, cytotoxins, drugs, growth factors, interferons, hormones, lipids, demineralized bone or bone morphogenetic proteins, cartilage inducing factors, oligonucleotides polymers, polysaccharides, polypeptides, protease inhibitors, vasoconstrictors or vasodilators, vitamins, minerals, stabilizers and the like. Also disclosed are methods of preparing and / or using the unsupplemented or supplemented fibrin sealant bandage.

A method of forming an array of viable cells is carried out by ink-jet printing a cellular composition containing said cells on a substrate. At least two different types of viable mammalian cells are printed on the substrate, the at least two different types of viable mammalian cells selected to together form a tissue. In some embodiments at least three or four different viable mammalian cells are printed on the substrate, the cells selected to together form a tissue. In some embodiments one of the viable mammalian cell types is a stem cell. In some embodiments the method further comprises printing at least one support compound on the substrate, the support compound selected to form a tissue together with said cells. In some embodiments the method further comprises printing at least one growth factor on the substrate, the growth factor selected to cause the cells to form a tissue.

Described herein are bioprinters comprising: one or more printer heads, wherein a printer head comprises a means for receiving and holding at least one cartridge, and wherein said cartridge comprises contents selected from one or more of: bio-ink and support material; a means for calibrating the position of at least one cartridge; and a means for dispensing the contents of at least one cartridge. Further described herein are methods for fabricating a tissue construct, comprising: a computer module receiving input of a visual representation of a desired tissue construct; a computer module generating a series of commands, wherein the commands are based on the visual representation and are readable by a bioprinter; a computer module providing the series of commands to a bioprinter; and the bioprinter depositing bio-ink and support material according to the commands to form a construct with a defined geometry.

A multi-layered microcapsule has an inner extracellular matrix and an outer shell. The inner extracellular matrix includes a first inner layer of biopolymer and a second intermediate layer of polymer that provides partial immune-protection and holds the first layer in place. The outer shell can form an exoskeleton to provide mechanical stability. Each of the individual layers can be varied to optimize mechanical stability, cell function, and immuno-protection.

This invention provides a fibrin sealant dressing, wherein said fibrin sealant may be supplemented with at least one composition selected from, for example, one or more regulatory compounds, antibody, antimicrobial compositions, analgesics, anticoagulants, antiproliferatives, antiinflammatory compounds, cytokines, cytotoxins, drugs, growth factors, interferons, hormones, lipids, demineralized bone or bone morphogenetic proteins, cartilage inducing factors, oligonucleotides polymers, polysaccharides, polypeptides, protease inhibitors, vasoconstrictors or vasodilators, vitamins, minerals, stabilizers and the like. Also disclosed are methods of preparing and / or using the unsupplemented or supplemented fibrin sealant dressing.

The present invention relates to a three-dimensional system, and compositions obtained therefrom, wherein individual layers of the system comprise channels divided longitudinally into two compartments by a centrally positioned membrane, and wherein each compartment can comprise a different cell type.

Methods of making a biologically active three-dimensional scaffold capable of supporting growth and differentiation of a cell are described. Biologically active three-dimensional scaffold made by the methods of the invention and an engineered tissue made from the scaffolds are described. Fibers of desired porosity can be obtained from non-structural ECM by lyophilization and / or electrospinning which can be useful for numerous tissue engineering applications requiring complex scaffolds, such as wound healing, artificial skin (burns), soft tissue replacement / repair and spinal cord injury.

Described herein are bioprinters comprising: one or more printer heads, wherein a printer head comprises a means for receiving and holding at least one cartridge, and wherein said cartridge comprises contents selected from one or more of: bio-ink, and support material; a UV light module for optionally exposing the contents of at least one cartridge to UV light; a means for calibrating the position of at least one cartridge; and a means for dispensing the contents of at least one cartridge. Also described herein are methods of using and bioprinting cartridges for such bioprinters.

Engineered animal skin, hide, and leather comprising a plurality of layers of collagen formed by cultured animal collagen-producing (e.g., skin) cells. Layers may be formed by elongate multicellular bodies comprising a plurality of cultured animal cells that are adhered and / or cohered to one another; wherein the elongate multicellular bodies are arranged to form a substantially planar layer for use in formation of engineered animal skin, hide, and leather. Further described herein are methods of forming engineered animal skin, hide, and leather utilizing said layers of animal collagen-producing cells.

Disclosed are living, three-dimensional tissue constructs for in vitro scientific and medical research, arrays thereof, and methods of making said tissues and arrays.

A multi-layered tissue construct includes: a first layer comprising a first hydrogel; and a second layer comprising a second hydrogel, wherein the first layer is connected to the second layer at a first transition zone and wherein at least one of the first layer and the second layer further comprises a component selected from the group consisting of cells and a bioactive substance. Another multi-layered tissue construct includes: a first layer comprising a first hydrogel; a second layer comprising cells of a first type, wherein the second layer is disposed on the first layer; and a third layer comprising a second hydrogel and optionally cells of the first type encapsulated in the second hydrogel, wherein the third layer is disposed on the second layer. Methods for producing these multi-layered tissue constructs are also disclosed.

The invention discloses a method for culturing normal human lung tissue and a lung cancer tissue organoid in an in-vitro manner. The method includes: acquiring fresh human-derived lung tissue cells, and performing collagen digestion on the fresh human-derived lung tissue cells to obtain single cells; culturing human lung tissue and the lung cancer tissue organoid under in-vitro 3D culture conditions; performing H&E staining to determining the structure and form, and using q-PCR to detect related gene expression; using immunofluorescent staining to authenticate cell sources and detect related protein expression. The invention further discloses a method for building a mouse animal model based on the organoid. The method for culturing the normal human lung tissue and the lung cancer tissue organoid and the method for building the mouse animal model have the advantages that the methods are significant to the building of large-scale and good-consistency human-derived in-situ lung cancer animal models, and a good basis and related application prospect are provided for the fundamental researches of lung cancer.

Described herein are bioprinters comprising: one or more printer heads, wherein a printer head comprises a means for receiving and holding at least one cartridge, and wherein said cartridge comprises contents selected from one or more of: bio-ink and support material; a means for calibrating the position of at least one cartridge; and a means for dispensing the contents of at least one cartridge. Further described herein are methods for fabricating a tissue construct, comprising: a computer module receiving input of a visual representation of a desired tissue construct; a computer module generating a series of commands, wherein the commands are based on the visual representation and are readable by a bioprinter; a computer module providing the series of commands to a bioprinter; and the bioprinter depositing bio-ink and support material according to the commands to form a construct with a defined geometry.

Three-dimensional tissue-like organization of cells by high cell-seeding-density culture termed as macromass culture is described. By macromass culture, cells can be made to organize themselves into a tissue-like form without the aid of a scaffold and three-dimensional macroscopic tissue-like constructs can be made wholly from cells. Tissue-like organization and macroscopic tissue-like constructs can be generated from fibroblastic cells of mesenchymal origin (at least), which can be either differentiated cells or multipotent adult stem cells. In this work, tissue-like organization and macroscopic tissue-like constructs have been generated from dermal fibroblasts, adipose stromal cells-derived osteogenic cells, chondrocytes, and from osteoblasts. The factor causing macroscopic tissue formation is large scale culture at high cell seeding density per unit area or three-dimensional space, that is, macromass culture done on a large scale. No scaffold or extraneous matrix is used for tissue generation, the tissues are of completely cellular origin. No other agents (except high cell-seeding-density) that aid in tissue formation such as tissue-inducing chemicals, tissue-inducing growth factors, substratum with special properties, rotational culture, etc, are employed for tissue formation. These tissue-like masses have the potential for use as tissue replacements in the human body. Tissue-like organization by high cell-seeding-density macromass culture can also be generated at the microscopic level.

The invention is a cell aggregation device comprising a hydrogel substrate having at least one, preferably a plurality, of cell-repellant compartments recessed into the uppermost surface. Each compartment is composed of an upper cell suspension seeding chamber having an open uppermost portion and a bottom portion, and one, or more than one, lower cell aggregation recess connected to the bottom portion of the upper cell suspension seeding chamber by a port. The diameter of the port may be fully contiguous with the walls of the chambers and walls of the recesses, or the diameter of the port may be more narrow than the walls of the chamber but fully contiguous with the walls of the recesses or more narrow than both the walls of the chamber and the walls of the recesses. The upper cell suspension seeding chambers are formed and positioned to funnel the cells into the lower cell aggregation recesses through gravitational force. The aggregation recesses are formed and positioned to promote cellular aggregation by coalescing cells into a finite region of minimum gravitational energy, increasing intercellular contact and minimizing or preventing cell adherence to the substrate. A device for encapsulating aggregates of live cells is provided. The device comprises (i) a biocompatible, bio-sustainable substrate having a cell-encapsulating face composed of one or more biocompatible, bio-sustainable, spaced-apart, cell-encapsulating compartments extending therefrom and (ii) a coating layer composed of a biocompatible, bio-sustainable polymer that completely surrounds the substrate and the cell-encapsulating compartments. A method for making the device is also provided.

Disclosed are engineered, living, three-dimensional connective tissue constructs comprising connective tissue cells. In some embodiments, the connective tissue cells are derived from multi-potent cells such as mesenchymal stem / stromal cells. In some embodiments, the cells are cohered to one another. In some embodiments, the multi-potent cells have been exposed to one or more differentiation signals to provide a living, three-dimensional connective tissue construct. In some embodiments, the constructs are substantially free of pre-formed scaffold at the time of use. Also disclosed are implants for engraftment, arrays of connective tissue constructs for in vitro experimentation, as well as methods of making the same.

Engineered animal skin, hide, and leather comprising a plurality of layers of collagen formed by cultured animal collagen-producing (e.g., skin) cells. Layers may be formed by elongate multicellular bodies comprising a plurality of cultured animal cells that are adhered and / or cohered to one another; wherein the elongate multicellular bodies are arranged to form a substantially planar layer for use in formation of engineered animal skin, hide, and leather. Further described herein are methods of forming engineered animal skin, hide, and leather utilizing said layers of animal collagen-producing cells.

A cell culture method prepares first cells that are adhesion-dependent cells and monolayered or multilayered cells on a culture surface of a culture substrate, seeds second cells that are adhesion-dependent cells and are magnetized by allowing to have magnetic particles on the first cells, induces the second cells to a predetermined position on the first cells by magnetic force, and cultures the first cells and the second cells in a cell arrangement obtained by the magnetic induction. According to this cell culture method, after a cell sheet was prepared individually, cells can be multilayered without changing a temperature, peeling the monolayered sheet and laminating the monolayered sheets.

A process and apparatus are provided for manufacturing complex parts and devices which utilize a CAD environment to design a part or device to be created (FIG. 1); Boolean, scaling, smoothing, mirroring, or other operations to modify the CAD design; a software interface to convert the CAD designed part (Data Process System) or device into a heterogeneous material and multi-part assembly model (Design Input Model) which can be used for multi-nozzle printing; and a multi-nozzle system to print the designed part or device using different, specialized nozzles (Tissue substitutes).