117 results about "Mature cell" patented technology

This disclosure provides a newly developed strategy and particular options for differentiating pluripotent stem cells into cells of the hepatocyte lineage. Many of the protocols are based on a strategy in which the cells are first differentiated into early germ layer cells, then into hepatocyte precursors, and then into mature cells. The cells obtained have morphological features and phenotypic markers characteristic of human adult hepatocytes. They also show evidence of cytochrome p450 enzyme activity, validating their utility for commercial applications such as drug screening, or use in the manufacture of medicaments and medical devices for clinical therapy.

The invention described herein provides a structure for growing isolated differentiable human mesenchymal cells, which includes a three-dimensional matrix of fibers. The matrix serves as an implantable scaffolding for delivery of differentiable human mesenchymal cells in tissue engineering. The invention further provides compositions that contain the three-dimensional matrix of fibers seeded with isolated differentiable human mesenchymal cells, wherein the matrix forms a supporting scaffold for growing the isolated differentiable human mesenchymal cells, and wherein the differentiable human mesenchymal cells differentiate into a mature cell phenotype. The invention further provides methods of preparing the implantable nanofiber matrix scaffolding seeded with differentiable human mesenchymal cells for use in tissue engineering.

This invention provides an isolated electrophysiologically immature cell or its derivative that has been modified to provide a mature electrophysiological phenotype and populations of these cells. Compositions containing these cells and populations of cells are also provided by this invention. These cells and compositions have therapeutic and diagnostic uses. Non-limiting therapeutic uses include regenerating cardiac tissue, improving cardiac function, restoring action potential of cardiac tissue; and treating or preventing cardiac malfunction. The cells and population of cells also can be used diagnostically to screen drug or other therapeutic candidate.

A production method for producing cell colony forming units in vitro from a mammalian tissue explant, the method comprises the steps of a) growing a piece of the mammalian tissue explant in a growth medium to obtain cell colony forming units from immature cells from the piece of explant, and b) harvesting cells from one or more of the cell colony forming units for use in Autologous Cell Implantation / transplantation methods. The mammalian tissue explant is selected from the group consisting of cartilage; bone such as, e.g., bone marrow; connective tissue; muscle tissue such as, e.g., smooth muscle tissue, heart tissue, liver tissue and skeletal muscle tissue; skin tissue such as, e.g., periosteum; mucosal tissue; brain tissue, pancreas tissue and blood vessels. In particularly, the mammalian tissue explant is cartilage, such as elastic, fibro, hyalin or articular hyalin cartilage. The cells obtained are suitable for use in autologous implantation / transplantation methods. In a specific embodiment, the cell obtained are chondroytes, especially for use in autologous chondrocyte implantation (ACI) methods.

The invention provides a biological artificial trachea and a preparation method thereof in combination with a 3D printing technology. Materials adopted for the biological artificial trachea simultaneously have flexibility and a certain mechanical strength, so that physical and mechanical properties of the trachea serving as a hollow organ in the chest are met. The artificial trachea is porous and loosened in structure and favorable for nutrient diffusion and ingrowth of autologous vessels. In addition, seed cells obtained by expansion of airway epithelial cells and cartilage cells of a receptor are printed on the trachea wall, and patients' rejection reaction to cells is avoided. Besides, the seed cells are composed of mature cells in terminal differentiation, the induced differentiation problem and the tumor formation risk caused by the use of stem cells can be avoided. The spatial distribution of the cells is accurately controllable in printing, and the restorative process is accelerated according to the histology regular distribution. Finally, a trachea support can be degraded in vivo, complications caused by the fact that foreign matter remains in vivo are reduced, and the risk of taking out the foreign matter through a second operation is avoided.

Undifferentiated primordial stem cells are manipulated to permit their long term growth in defined media lacking serum and feeder layer cells by shifting the apoptotic balance of the cells, through increasing the activity of Bcl-2 family anti-apoptotic proteins or decreasing the activity of Bcl-2 family pro-apoptotic proteins. In some embodiments of the invention, the Bcl family protein is Bcl-2. The ES cells sustain the characteristics of undifferentiated, pluripotent stem cells during long-term serum- and feeder layer cell-free conditions, including the ability to be expanded in vitro, but maintain their potential to differentiate into mature cell types.

Vascular scaffolds and methods of fabricating the same are disclosed for tissue engineering of vascular constructs. By combining electrospun matrices with cell sheet technologies, vascular constructs with more mature cell layers can be obtained for reconstruction of blood vessels, heart valves and the like. A engineered smooth muscle cell sheet, wrapped around an electrospun vascular scaffold, is able to provide a mature SMC layer that expresses strong cell-to-cell junction markers and contractile proteins. In addition, preconditioning of the cell sheet covered vascular scaffold maintained cell viability and infiltration into the scaffold.

The invention discloses a preparation method of antigen-specific cytotoxicity T lymphocytes. The method comprises the following steps: 1, constructing a recombinant vector carrying tumor antigen gene; 2, separating and culturing DC cells; 3, transducing immature DC cells in step 2 by using the recombinant vector in step 1; 4, inducing the immature DC cells in step 3 through cytokines to form mature DC cells; and 5, coculturing the mature DC cells in step 4 and T lymphocytes to obtain the antigen-specific cytotoxicity T lymphocytes. The method preferably selects a lentivirus mixed vector realizing transduction to be used in the expression of three wide spectrum tumor antigens comprising CEA, MAGE-A3 and MUC1 to obtain activated and proliferated tumor antigen-specific T cells, and has a wide application prospect.

We describe an improved method for generating sizable numbers of mature dendritic cells from nonproliferating progenitors in human blood. The first step or “priming” phase is a culture of T cell depleted mononuclear cells in medium supplemented with GM-CSF and IL-4 to produce immature dendritic cells. The second step or “differentiation” phase requires the exposure to dendritic cell maturation factor such as monocyte conditioned medium. Using this two-step approach, substantial yields are obtained. The dendritic cells derive from this method have all the features of mature cells. They include a stellate cell shape, nonadherence to plastic, and very strong T cell stimulatory activity. The mature dendritic cells produced according to this invention are useful for activating T cells.

Disclosed are a composition for introducing the osteogenic differentiation of human embryonic stem cells and a method for differentiating human embryonic stem cells into an osteoblastic lineage by inhibiting the mTOR signaling pathway. When cultured in the presence of an inhibitor of the mTOR signaling pathway, human embryonic stem cells are effectively induced to differentiate into an osteoblastic lineage. The osteogenic differentiation of human embryonic stem cells using the method and the composition is useful in examining the development and differentiation mechanism of osteoblasts and the cause of metabolic bone diseases, including osteoporosis. In addition, the method and the composition can be applied to the development of osteogenic differentiation techniques for generating clinically useful, terminally differentiated mature cells or progenitor cells.

The invention relates to a method for carrying out the expansion of multipotent stem cells ex vivo. Moreover, the invention relates to a two-stage method for carrying out the expansion and differentiation of multipotent stem cells ex vivo, in which it is possible for the stem cells to be gene transfected during the first stage, i.e. during the expansion phase. In this phase, the differentiation of the multipotent stem cells takes place optionally in cells of the hematopoietic, endothelial or mesenchymal cell lineage. Stem and progenitor cells as well as mature cells of the hematopoietic, endothelial and mesenchymal cell lineage, which are obtained in this way, can be used, among other things, for the prophylaxis, diagnosis and therapy of human deseases and for tissue engineering.

The invention discloses a modified enhanced type DC-CIK cell targeting immune cell mass, and a preparation method and application thereof. A method for preparing a DC cell comprises the following steps: performing isolated culture on monocyte, thereby acquiring an immature DC cell; and performing tumor antigen load and induced differentiation culture on the immature DC cell, thereby acquiring a mature DC cell, wherein the monocyte is acquired by separating from a peripheral blood mononuclear cell of a sample. The method for preparing CIK comprises the following steps: performing in vitro differentiation culture on a CIK cell, thereby acquiring an immature CIK cell; preparing a modifying recombined slow virus for performing in vitro modification on the CIK cell; and co-culturing the modified CIK cell and the DC cell, thereby acquiring the mature modified CIK cell (Triumph-DC-CIK), so as to be used for achieving a better tumor immunotherapy effect.

A tissue structure for enabling comprehensive understanding of gene patterns of mature cells and a method of preparing the tissue structure are provided. A tissue structure is obtained by co-culturing an endodermal, ectodermal, or mesodermal cell derived from a stem cell and at least one cell and / or factor selected from the group consisting of a vascular cell, a mesenchymal cell, a factor secreted by a vascular cell, a factor secreted by a mesenchymal cell, and a factor secreted when both a vascular cell and a mesenchymal cell exist. A value obtained by assay of a plurality of functions using a Pearson product-moment correlation coefficient is closer to a value of a cell or biological tissue sampled from an adult than a value of a cell or biological tissue sampled from a fetus.

This invention relates to a method of delivering light energy to biological tissues for the acceleration of healing of damaged or diseased tissues or regeneration of such tissues. More particularly, the present invention relates to applying various wavelengths of light to articular and non-articular joints, alone, or in conjunction with techniques for restoring or regenerating cartilage, ligament and / or tendons whether in-vitro, in-situ or in-vivo. The present invention also extends to application of photo-irradiation to stimulate enhanced proliferation and site-dependent differentiation of stem cells into mature cells and to stimulate full functioning of mature cells involved in tissue repairs for regeneration of tendons, ligaments, cartilage, bone or muscle, depending on the type of tissue with which the stem cells come into contact.

The present invention is a method for clinic hematopoietic stem cells transplantation by extracting original mesenchymal and hematopoietic stem / progenitor cells from health post-caesarean birth placenta tissue. Each placenta can extract 3.8 + / - 0.7 X 10<9> cells having nucleus, wherein original mesenchymal and hematopoietic stem / progenitor cells are 1.8 + / - 0.2 X10<7>, cell viability is more than 95 percent, microorganism contamination rate is less than 6 percent. The method uses counter monoclonal antibody method to eliminate 90 to 95 percent contaminated mature cell having nucleus single cell in mother blood, reducing immunological rejection reaction (GVHD) after transplanting. The finished product of the method can be used for clinic treating hematopoietic stem cells transplantation due to marrow injury, can be used for applying to placenta tissue supplying person or other patient having various hematopoietic stem cells transplantation indications, and can also combine with various cell growth factors in clinic to develop cell repair and gene treatment. The present invention supplies another cell source for hematopoietic stem cells transplantation, promotes the development of cells transplantation and tissue repairing, also supplies new research direction for early tissue cell differentiation.

The invention discloses a DC cell and a targeting immune cell population, and a preparation method and application thereof. The preparation method of DC cells includes the steps of: conducting first induced differentiation culture on mononuclear cells in order to obtain immature DC cells; transfecting the immature DC cells by using HSV-2 antigen mRNA, so as to obtain transfected immature DC cells; and conducting second induced differentiation culture on the transfected immature DC cells, so as to obtain mature DC cells, wherein the mononuclear cells are isolated from mononuclear cells of peripheral blood of a sample. The method can prepare DC cells safely and efficiently, has a low cost and high efficiency, and can effectively solve the problem of cytotoxicity. The method can be effectively used for immune cell treatment or stimulation of lymphocytes derived from a same patient to produce a targeting cell population containing a large amount of CTL cells, and then the cell population can be used for virus infection immunotherapy to reach better effect.

The invention discloses a fat particle separation and purification device and a fat particle separation and purification method thereof. The fat particle separation and purification device comprises acover and a barrel, the top of the barrel is covered by the cover, so that a fully closed fat storage barrel is formed, the cover is provided with a negative pressure connector and a fat suction connector, at least one filter is arranged inside the barrel, the at least one filter in the barrel divides the barrel into a plurality of separation chambers from the top down, a rotary shaft is arrangedat the center of the cover, a separating scraper is arranged on the rotary shaft, close to the upper surface of each filter, a plurality of rows of fine stainless steel wires which are longitudinallyarranged are mounted on the rotary shaft in the sample collection chamber, and the plurality of rows of fine stainless steel wires are distributed around the rotary shaft to form separating brackets.The fat particle separation and purification device disclosed by the invention can non-centrifugally automatically separate and remove fibrous tissues, disrupted mature cell oil drops and tumescent fluid in fat mixed liquid under a fully closed state, and can screen different grain sizes of fat particles layer by layer.

The present invention, in some embodiments, describes methods for selecting and enriching specific types of stem cells and / or progenitor cells from a sample containing at least two different types of stem cells and / or progenitor cells. In one embodiment, the method involves selecting and enriching cancer stem cells and / or cancer progenitor cells. In other embodiments, the invention involves improved methods for purging cancer cells from autologous or allogenic transplants prior to reinjection into a patient. In other embodiments, the invention describes improved methods to screen for the efficacy of drug candidate for affecting the function and / or viability of a specific type of stem cell or progenitor cell, for example a cancer stem cell. In this context, the inventive method can involve screening the effectiveness of chemotherapeutic agents in completely eliminating all cancer cells (i.e. mature cancer cells and cancer stem / progenitor cells). In yet other embodiments, the invention provides methods for the identification of a particular type or sub-population of stem cells or progenitor cells from a mixed cell-type population. In one embodiment of such a method, the invention describes a method for the early detection of metastatic cancer from simple tests on blood or bone marrow. Stem cell culture methods are also disclosed which utilize cell lysates to preferentially induce either asymmetric or symmetric division of the stem cells. The invention also provides, in some embodiments, novel stem cell suspensions that include, in some instances enriched suspensions of particular types or sub-populations of stem cells or progenitor cells, for example cancer stem cells. In other embodiments, the invention provides a suspension of stem cells that is substantially free of all stem cells of a particular type or sub-population and, optionally, also substantially free of all mature cells. In one example of such an embodiment, the invention provides a suspension of stem cells that is substantially free of all cancer cells (i.e. mature cancer cells as well as cancer stem cells and cancer progenitor cells).

The invention provides umbilical cord mesenchymal stem cells for treating lung diseases and a preparation method thereof. The umbilical cord mesenchymal stem cells provided by the invention can be used for preparing drugs for preventing or treating pneumoconiosis injury and fibrosis. The drugs are high in safety and good in intervention and treatment effect on corresponding diseases when being applied to lung disease treatment, especially pneumoconiosis lung injury and fibrosis thereof, quantitative production can be achieved through mature cell culture technical means, and the medicine has bright application prospects.

The invention relates to an in-vitro amplification culture method and application of hepatocytes. According to the invention, a culture system for reprogramming human hepatocytes into proliferative intermediate state cells between mature hepatocytes and liver precursor cells is established, and the liver reproductive capacity of the human hepatocytes is verified in animal bodies. According to themethod, exogenous genes do not need to be introduced into the hepatocytes; amplification of the hepatocytes can be achieved through conventional culture; the obtained hepatocytes can be subjected to passage; and maturation culture can be further carried out to obtain functional human liver maturation cells.

The invention discloses a DC cell based on SP17 antigen, a targeting immune cell population, a preparation method and applications thereof. The preparation method of DC cell comprises the following steps: subjecting mononuclear cells to first induced differentiation culture to obtain immature DC cells; transfecting the immature DC cells by SP17 antigen mRNA to obtain transfected immature DC cells; subjecting the transfected immature DC cell to second induced differentiation culture to obtain mature DC cells; wherein the mononuclear cells are separated from peripheral blood mononuclear cells of a sample. Through the provided preparation method, DC cells can be obtained efficiently and safely. Moreover, the provided preparation method has the advantages of low cost and high efficiency, and can effectively solve the problem of cell toxicity. The DC cells can be applied to cell immunotherapy or used to trigger the lymph cells from the same patient to generate targeting immune cell population rich in CTL cells so as to achieve a better effect in tumor immunotherapy.

The present invention provides a biological composite artificial trachea and its preparation method in combination with 3D printing technology. The material used in the present invention has both flexibility and certain mechanical strength, which meets the physical and mechanical properties of the trachea as a hollow organ in the thoracic cavity; secondly, The artificial trachea of ​​the present invention has a porous and loose structure, which is conducive to the diffusion of nutrients and the growth of autologous blood vessels; in addition, the present invention prints the seed cells obtained from the expansion of the recipient's own airway epithelial cells and chondrocytes in the trachea wall, The patient's rejection of the cells is avoided; moreover, the seed cells are composed of terminally differentiated mature cells, which avoids the problem of inducing differentiation and the risk of tumor formation caused by the use of stem cells; at the same time, the spatial distribution of the cells in the present invention is The printing is precise and controllable, and the distribution according to histological laws accelerates the repair process; finally, the tracheal stent can be degraded in the body, reducing the complications caused by foreign body residues in the body, and avoiding the risk of secondary surgical removal.

Improved colony forming cell (CFC) assays are described. The improved assay comprises modifications to the standard CFC assay that enable analysis of temporal, real-time changes in antigen expression during colony development without need to fixate or destroy the culture. The improved assay is applicable to hematopoietic CFC assays as well as to CFC assays for other cells, such as neural cells and mammary cells. In one embodiment, the invention comprises adding a detection reagent, most likely a fluorescently labeled antibody, that is specific for antigens (most likely a cell-surface antigen) expressed on progenitors or on specific mature cell types to a culture at the start or during culture. The invention also comprises modifications to the culture medium and cell preparations used in standard CFC assays to selectively promote the development of one colony type while preventing or suppressing the development of other colony types.

Undifferentiated primordial stem cells are manipulated to permit their long term growth in defined media lacking serum and feeder layer cells by shifting the apoptotic balance of the cells, through increasing the activity of Bcl-2 family anti-apoptotic proteins or decreasing the activity of Bcl-2 family pro-apoptotic proteins. In some embodiments of the invention, the Bcl family protein is Bcl-2. The ES cells sustain the characteristics of undifferentiated, pluripotent stem cells during long-term serum- and feeder layer cell-free conditions, including the ability to be expanded in vitro, but maintain their potential to differentiate into mature cell types.

The invention discloses a preparation kit and a preparation method of adherent mononuclear cells, immature DCs (Dendritic Cells) and mature DCs, and an application of the mature DCs prepared by the method in preparing an anti-tumor cell preparation, and belongs to the biological technical field. According to the preparation kit, the preparation method and the application of the mature DCs, which are disclosed by the invention, the adherent mononuclear cells are prepared from CD14 monoclonal antibodies through a culture bottle dish made of a polystyrene, polyvinyl chloride or polyethylene material; the immature DCs are prepared from GM-CSF (granulocyte-macrophage colony-stimulating factor), IL-4 (interleukin-4) and IFN (interferon)-alpha; the mature DCs are prepared from TNF (tumor necrosis factor)-alpha and OK-432. The method disclosed by the invention is capable of increasing the adherence ability of the adherent mononuclear cells and increasing the number of the adherent cells, and thus being advantageous for reducing the influence of impurity cells on the transformation of the adherent mononuclear cells into the immature DCs; besides, the method is capable of increasing the yield of the immature DCs; the method is capable of obtaining mature DCs which are higher in maturity, and better in cell migration ability and IL-12p70 secretion ability.

Provided are age-modified cells and method for making age modified cells by reducing or increasing the level of genomic nucleic acid methylation in the cells. The aging and / or maturation process can be accelerated or reduced and controlled for young, aged, mature and / or immature cells, such as a somatic cell, a stem cell, a stem cell-derived somatic cell, including an induced pluripotent stem cell-derived cell, by reducing or increasing the level of genomic nucleic acid methylation in the cells. Methods described by the present disclosure can produce age-appropriate cells from a somatic cell or a stem cell, such as an old cell, young cell, immature cell, and / or a mature cell. Such age-modified cells constitute model systems for the study of late-onset diseases and / or disorders.

The present invention relates generally to the field of tissue engineering and more particularly to a method for generating tissue suitable for use in tissue replacement and / or tissue rejuvenation therapy and / or as a source of cell-derived therapeutic or diagnostic agents including proteins and hormones. Even more particularly, the present invention contemplates the use of cell fusion techniques involving single cell, mini-bulk or macro-bulk cell fusion to generate tissue or cells useful for tissue replacement and / or tissue rejuvenation therapy or a range of organs or areas of the body. The resulting tissue or cells may also secrete or generate a range of cytokines, enzymes, hormones and the like which have improved or more efficacious properties relative to analogous molecules produced from non-fused cells. The present invention further provides an apparatus having aspects controlled by data processing means which facilitates the fusion of a pair of cells. Of the pair of cells, at least one of the cells in the pair may be a mature cell or is capable of differentiating or developing into a mature cell. The subject invention further provides isolated molecules such as cytokines, receptors, antibodies, hormones, heat shock proteins, enzymes, and glycoproteins such as mucins, lectins and heparan sulfates derived from fused cells. These molecules may be characterized by having altered glycosylation patterns, altered post-translational modifications, greater activity, being more efficacious or being more stable relative to analogous molecules from non-fused cells. The present invention further provides novel cell fusates or cell hybrids having a pattern of cell surface markers unique relative to the at least two cells which fuse together to generate the cell. These cell markers are useful in selecting particular cell hybrids and as proprietary tags.