303 results about "Directed differentiation" patented technology

Stem cells, including mammalian, and particularly primate primordial stem cells (pPSCs) such as human embryonic stem cells (hESCs), hold great promise for restoring cell, tissue, and organ function. However, cultivation of stem cells, particularly undifferentiated hESCs, in serum-free, feeder-free, and conditioned-medium-free conditions remains crucial for large-scale, uniform production of pluripotent cells for cell-based therapies, as well as for controlling conditions for efficiently directing their lineage-specific differentiation. This instant invention is based on the discovery of the formulation of minimal essential components necessary for maintaining the long-term growth of pPSCs, particularly undifferentiated hESCs. Basic fibroblast growth factor (bFGF), insulin, ascorbic acid, and laminin were identified to be both sufficient and necessary for maintaining hESCs in a healthy self-renewing undifferentiated state capable of both prolonged propagation and then directed differentiation. Having discerned these minimal molecular requirements, conditions that would permit the substitution of poorly-characterized and unspecified biological additives and substrates were derived and optimized with entirely defined constituents, providing a “biologics”-free (i.e., animal-, feeder-, serum-, and conditioned-medium-free) system for the efficient long-term cultivation of pPSCs, particularly pluripotent hESCs. Such culture systems allow the derivation and large-scale production of stem cells such as pPSCs, particularly pluripotent hESCs, in optimal yet well-defined biologics-free culture conditions from which they can be efficiently directed towards a lineage-specific differentiated fate in vitro, and thus are important, for instance, in connection with clinical applications based on stem cell therapy and in drug discovery processes.

Stem cells, including mammalian, and particularly primate primordial stem cells (pPSCs) such as human embryonic stem cells (hESCs), hold great promise for restoring cell, tissue, and organ function. However, cultivation of stem cells, particularly undifferentiated hESCs, in serum-free, feeder-free, and conditioned-medium-free conditions remains crucial for large-scale, uniform production of pluripotent cells for cell-based therapies, as well as for controlling conditions for efficiently directing their lineage-specific differentiation. This instant invention is based on the discovery of the formulation of minimal essential components necessary for maintaining the long-term growth of pPSCs, particularly undifferentiated hESCs. Basic fibroblast growth factor (bFGF), insulin, ascorbic acid, and laminin were identified to be both sufficient and necessary for maintaining hESCs in a healthy self-renewing undifferentiated state capable of both prolonged propagation and then directed differentiation. Having discerned these minimal molecular requirements, conditions that would permit the substitution of poorly-characterized and unspecified biological additives and substrates were derived and optimized with entirely defined constituents, providing a “biologics”-free (i.e., animal-, feeder-, serum-, and conditioned-medium-free) system for the efficient long-term cultivation of pPSCs, particularly pluripotent hESCs. Such culture systems allow the derivation and large-scale production of stem cells such as pPSCs, particularly pluripotent hESCs, in optimal yet well-defined biologics-free culture conditions from which they can be efficiently directed towards a lineage-specific differentiated fate in vitro, and thus are important, for instance, in connection with clinical applications based on stem cell therapy and in drug discovery processes.

The present invention provides methods for the directed differentiation of embryonic stem cells along the endodermal lineage, especially the pancreatic lineage.

Methods are described for mapping a pathway of differentiation of a population of embryonic cells which includes exposing the cells to an exogenous factor and measuring gene expression products that are characteristic of a particular cell type or lineage. Directing differentiation of human embryonic cells relies on dissociated embryoid bodies which are then exposed to one or more exogenous factors to enrich a culture for a particular cell type. The differentiated cells may be used for treating a medical condition in a human. Kits for determining differentiation pathways and screening exogenous factors for their utility in differentiation are provided.

The present invention provides a preparation of undifferentiated embryonic stem (ES) cells sustainable for a prolonged period in an undifferentiated state which will undergo stem cell renewal or somatic differentiation. Preferably the cells are capable of somatic differentiation in vitro and are inclined to differentiate away from an extraembryonic lineage. The present invention also provides method of culturing embryonic stem (ES) cells to improve stem cell maintenance and persistence in culture. The method also provides a culture of ES cells prepared by the method as well as differentiated cells derived from the embryonic cells resulting from directed differentiation procedures provided by the present invention.

The present invention concerns RPE cells obtainable by directed differentiation from stem cell, particularly, human stem cells. It has been specifically found that culturing stem cells in the presence of one or more member of the TGFβ superfamily, such as Activin A) induced directed differentiation into mature and functional RPE cells. This was evidenced by the expression of markers specific to mature RPE cells, including MiTF-A, RPE65 or Bestrophin). In accordance with one particular embodiment, the cells are a priori cultured with nicotinamide (NA) which was found to augment the cells' response to the inductive effect of the one or more member of the TGFβ superfamily. The invention also provides methods of performing the directed differentiation, as well as methods for use of the resulting RPE cells.

The present invention relates generally to the field of cell biology of stem cells, more specifically the directed differentiation of pluripotent or multipotent stem cells, including human embryonic stem cells (hESC), somatic stem cells, and induced human pluripotent stem cells (hiPSC) using novel culture conditions. Specifically, methods are provided for obtaining neural tissue, floor plate cells, and placode including induction of neural plate development in hESCs for obtaining midbrain dopamine (DA) neurons, motorneurons, and sensory neurons. Further, neural plate tissue obtained using methods of the present inventions are contemplated for use in co-cultures with other tissues as inducers for shifting differentiation pathways, i.e. patterning.

The invention discloses a bionic three-dimensional tissue engineering scaffold, which is formed by high molecular fibrous membrane-loaded active growth factors, or active growth factors loaded on a composite scaffold formed by a high molecular fibrous membrane and a macropore spongy layer. With the adoption of the bionic three-dimensional tissue engineering scaffold, the problem that the concentration of active molecules loaded with an emulsion electricity texture fibrous membrane is low; the emulsion electricity texture fibrous membrane is combined with a macropore spongy or a mixed electricity texture process, so that the load rate of the active factors can be greatly improved; parts of factors are retained in the fiber through an emulsion electricity texture core-shell structure, so that the effective control of releasing time is realized, and a repairing process is monitored for a long time; and the introduction of active molecules in the scaffold plays guiding and promoting functions for proliferating regenerative cells, directionally differentiating, migrating and adhering cells, and capturing stem cells to introduce regenerative functions of newly born tissues, so that a new path is provided for development of regenerative medicine industries.

A modified P. aeruginosa type III secretion system has been developed that efficiently delivers selected proteins into a host cell. In one example, a functional nuclear Cre Recombinase is injected into embryonic stem (ES) cells and can be used to induce pluripotent stem (iPS) cells. This method of in vitro lineage directed differentiation prevents insertional mutagenesis and provides a route to selected stem cell renewal and cell-based therapies.

The invention relates to a tissue engineering bracket material capable of physically embedding active substances and a preparation method thereof. The tissue engineering bracket material capable of physically embedding the active substances is prepared from 100 weight parts of biodegradable macromolecular substance and 0 to 20 weight parts of active substances, wherein the weight part of the active substances is not equal to zero; 100 weight parts of biodegradable macromolecular substance consists of 50 to 100 weight parts of synthetic macromolecular substance and 0 to 50 weight parts of natural macromolecular substance; the macromolecular substances and the active substances form a micromolecular nanofiber membrane or a composite macromolecular nanofiber membrane; and the active substances are bonded on the surface of the fiber membrane uniformly. The tissue engineering bracket material capable of physically embedding the active substances has high biocompatibility and sufficient mechanical strength, can be biodegraded, is suitable for proliferation and directed differentiation of stem cells, and has the functions of promoting cell migration adhesion and capturing the stem cells so as to induce regeneration of tissues such as bones, cartilages, nerves, skin and the like.

The invention provides icariin application in inducing stem cell in vitro orientation differentiates to several single type cells. The stem cell said includes embryonic stem cell, nerve stem cell and marrow mesenchyme stem cell. The single type cell includes nerve cell, bone cell, islet cells and endothelial cell. This invention also relates to application of the single type cell in preparing medicine of stem cell transplant curing nerve degenerative diseases, and its application in preparing cell differentiation agent that used to repair recovery injured nerve tissue, and its application in high efficiency drug effect screening model rebuilding and initial screening and estimating by using the model. The new applications of the icariin is extended in this invention, the fact of icariin contains pharmacy activity clarified, so it provides material basis for Chinese traditional medicine prevention and cure effect. The clarifying of drug effect mechanism provides reference for new medicine Chinese metical modern development with self-owned intellectual property right.

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.

Provided are cortical interneurons and other neuronal cells and in vitro methods for producing such cortical interneurons and other neuronal cells by the directed differentiation of stem cells and neuronal progenitor cells. The present disclosure relates to novel methods of in vitro differentiation of stem cells and neural progenitor cells to produce several type neuronal cells and their precursor cells, including cortical interneurons, hypothalamic neurons and pre-optic cholinergic neurons. The present disclose describes the derivation of these cells via inhibiting SMAD and Wnt signaling pathways and activating SHH signaling pathway. The present disclosure relates to the novel discovery that the timing and duration of SHH activation can be harnessed to direct controlled differentiation of neural progenitor cells into either cortical interneurons, hypothalamic neurons or pre-optic cholinergic neurons. The present disclosure also relates to compositions of cortical interneurons, hypothalamic neurons or pre-optic cholinergic neurons, and their precursors, that are highly enriched and can be used in variety of application. These cells can be used therapeutically to treat neurodegenerative and neuropsychiatric disorders, and can be used for disease modeling and drug screening.

The invention provides a method for directionally differentiating a multipotential stem cell in vitro into a myocardial cell and provides a culture medium for establishing myocardial cell differentiation by means of induction in vitro under a three-dimensional suspension condition. The culture medium comprises a differential medium for induced differentiation of a multipotential stem cell into a precursor cell of mesoblast, a differential medium for differentiating the precursor cell of mesoblast into the myocardial cell and a long-term maintaining culture medium of the myocardial cell. The method provided by the invention is simple and reliable, low in cost, stable and efficient and high in safety. By adopting a suspension culture system, high-quality myocardial cells can be industrially produced without any subsequent screening and purifying steps, can be directly used for application demands of scientific research of heart development, cell therapy of heart diseases, transplanting due to cardiac trauma and drug screening, and has inestimable scientific, social and economical benefits.

The invention provides a tissue engineering scaffold material of a chemical bonding active material and a preparation method of the tissue engineering scaffold material. The tissue engineering scaffold material of the chemical bonding active material is prepared by a biodegradable macromolecular substance and an active material, wherein the macromolecular substance is counted by 100 parts by weight, and the active material is 0-15 part(s) by weight but is not zero; the 100 parts by weight of the macromolecular substance consists of 50-100 parts by weight of the synthesized macromolecular substance and 0-50 part(s) by weight of natural macromolecular substance; and a macromolecular nano-fiber film or a composite macromolecular nano-fiber film is formed by the macromolecular substance by means of electrostatic spinning, and the active material is evenly bonded on the surface of the fiber film. The tissue engineering scaffold material of the chemical bonding active material provided by the invention is good in biocompatibility, enough in mechanical strength, biodegradable, and suitable for stem cell proliferation and directed differentiation, and has the functions of cell migration adhesion promotion and stem cell capture to induce the regeneration of tissues such as bone, cartilage, nerve, skin, etc.

Disclosed herein are methods for generating SC-β cells using chemically defined, completely serum free media, and isolated populations of SC-β cells for use in various applications, such as cell therapy.

The present invention concerns RPE cells obtainable by directed differentiation from stem cell, particularly, human stem cells. It has been specifically found that culturing stem cells in the presence of one or more member of the TGFβ superfamily, such as Activin A) induced directed differentiation into mature and functional RPE cells. This was evidenced by the expression of markers specific to mature RPE cells, including MiTF-A, RPE65 or Bestrophin). In accordance with one particular embodiment, the cells are a priori cultured with nicotinamide (NA) which was found to augment the cells' response to the inductive effect of the one or more member of the TGFβ superfamily. The invention also provides methods of performing the directed differentiation, as well as methods for use of the resulting RPE cells.

Methods of efficiently converting primate pluripotent stem cells to GABA neurons or cholinergic neurons, as well as applications thereof, are disclosed.

The generation of complex organ tissues from human embryonic and pluripotent stem cells (PSCs) remains a major challenge for translational studies. It is shown that PSCs can be directed to differentiate into intestinal tissue in vitro by modulating the combinatorial activities of several signaling pathways in a step-wise fashion, effectively recapitulating in vivo fetal intestinal development. The resulting intestinal “organoids” were three-dimensional structures consisting of a polarized, columnar epithelium surrounded by mesenchyme that included a smooth muscle-like layer. The epithelium was patterned into crypt-like SOX9-positive proliferative zones and villus-like structures with all of the major functional cell types of the intestine. The culture system is used to demonstrate that expression of NEUROG3, a pro-endocrine transcription factor mutated in enteric anendocrinosis is sufficient to promote differentiation towards the enteroendocrine cell lineage. In conclusion, PSC-derived human intestinal tissue should allow for unprecedented studies of human intestinal development, homeostasis and disease.

The present invention relates to the induction of differentiation in stem cells to cardiomyocytes and factors such as prostaglandin alone or in combination with other factors including essential minerals selected from the group including transferrin and selenium, small molecules selected from the group including a p38 MAPK inhibitor such as SB203580 and protein growth factors of the FGF, IGF and BMP families such as but not limited to IGF1, FGF2, BMP2, BMP4 and BMP6. and insulin that influence the process of differentiation to cardiomyocytes. Media that is appropriate for the induction of differentiation of cardiomyocytes from stem cells is also provided wherein the media contains these factors. The use of cardiomyocytes and cardiac progenitors produced by the directed differentiation in transplantation and screening for cardiac compounds is also provided.

The invention discloses a human mesenchymal stem cell adipogenesis inducing and differentiating culture medium and a preparation method thereof. The human mesenchymal stem cell adipogenesis inducing and differentiating culture medium is produced by the following components of an alpha-MEM / HG-DMEM culture medium, 5-50% of percent by volume of fetal calf serum, 0.5-10% of percent by volume of glutamine, 100-400 [mu]M volume of indomethacin, insulin with the concentration of 0.1-20 [mu]g / ml, 10-200 [mu]M volume of 1-methyl-3-isobutyl xanthine, 10-200 nM volume of dexamethasone and 0.1-20 [mu]M volume of spermine. The preparation method includes the following steps that a culture dish is cleaned; the culture medium is provided, and the alpha-MEM / HG-DMEM culture medium is prepared in the culture dish according to a formula; materials are mixed; and mixed liquor is filtered. Multiple histologic origin human mesenchymal stem cells including human mesenchymal stem cells, umbilical cord mesenchymal stem cells and adipose tissue-derived stromal cells are induced to the directional differentiation of adipogenesis cells; differentiating and inducing time of the human mesenchymal stem cells isshortened, the preparation method is convenient, and the differentiating and the inducing of human mesenchymal stem cell adipogenesis can be achieved stably and efficiently.

The generation of complex organ tissues from human embryonic and pluripotent stem cells (PSCs) remains a major challenge for translational studies. It is shown that PSCs can be directed to differentiate into intestinal tissue in vitro by modulating the combinatorial activities of several signaling pathways in a step-wise fashion, effectively recapitulating in vivo fetal intestinal al development. The resulting intestinal “organoids” were three-dimensional structures consisting of a polarized, columnar epithelium surrounded by mesenchyme that included a smooth muscle-like layer. The epithelium was patterned into crypt-like SOX9-positive proliferative zones and villus-like structures with all of the major functional cell types of the intestine. The culture system is used to demonstrate that expression of NEUROG3, a pro-endocrine transcription factor mutated in enteric anendocrinosis is sufficient to promote differentiation towards the enteroendocrine cell lineage. In conclusion, PSC-derived human intestinal tissue should allow for unprecedented studies of human intestinal development, homeostasis and disease.

The present invention relates to the use of 3D culturing systems for the derivation of hepatocyte-like cells from human pluripotent stem cells (hPS). In particular, the invention concerns the directed differentiation and maturation of human pluripotent stem cells into hepatocyte like cells in 3D hollow fiber capillary bioreactors.

The invention relates to a preparation method of a specific extracellular matrix ECM of periodontium. The preparation method comprises the following steps: 1) separating and culturing a human periodontal ligament cell; and 2) preparing the specific extracellular matrix ECM of periodontium; applying the specific extracellular matrix ECM of periodontium and applying the specific extracellular matrix ECM of periodontium as a human periodontal ligament stem cell in in-vitro amplification; and promoting directional differentiation of the ECM. The preparation method is simple and can remarkably promote in-vitro amplification and directional differentiation of the periodontal ligament stem cell.

The invention provides an umbilical cord mesenchymal stem cell injection with an anti-aging function and a preparation method thereof. The injection comprises the following components: umbilical cord mesenchymal stem cell (5*10<5> to 10*10<5> / mL), 2 wt% of human albumin, and 98 wt% of compound electrolyte solution. The produced umbilical cord mesenchymal stem cells can rapidly repair and replace damaged cells and tissues, and have the characteristics of amplification and directional cell differentiation; and after the stem cells receive the signals from damaged parts, the stem cells will move to the damage parts, amplify in the damage parts, carry out induced differentiation, promote the self recovery function of stem cells, rapidly repair and replace damaged cells and tissues, rapidly secrete cell factors, growth factors, and tissue related factors in time, thus improve the blood circulation, enhance the metabolism functions, improve the tissue repairing function and immunity, and finally achieve the anti-aging goal.

The invention relates to the technical field of biomedical engineering and particularly relates to a long-term in-vitro culture and directional differentiation system and method for a liver stem cell. The long-term in-vitro culture and directional differentiation system comprises an amplification culture medium with specific chemical components, and a differential medium with specific chemical components, wherein the amplification culture medium is used for carrying out in-vitro culture of a mouse or human liver stem cell, and the differential medium is used for carrying out induced differentiation on the mouse or human liver stem cell to form a matured liver cell. By using the long-term in-vitro culture and directional differentiation system and method, a selectively-amplified liver stem cell in mother cells of the liver can be obtained from a mouse embryonic liver tissue or through human multipotential stem cell differentiation, the liver stem cell can be cultured for more than 20 generations under such a condition, and the stable molecular phenotype of the liver stem cell is maintained. By using the long-term in-vitro culture and directional differentiation system and method, the cultured mouse or human liver stem cell can be further subjected to induced differentiation to form the matured liver cell with functions of secreting albumin, metabolizing urea and the like.

Disclosed herein are methods for generating SC-β cells using chemically defined, completely serum free media, and isolated populations of SC-β cells for use in various applications, such as cell therapy.

The invention relates to an extracellular matrix gel model used for researching development and differentiation of embryonic stem cells, belonging to the research field of tissue engineering and regenerative medicine, and mainly solving the problem that no ideal model exists for the research of differentiation and development of embryonic stem cells in vitro. We the extracellular matrix gel adopt as the in vitro three dimensional model of the differentiation and development of embryonic stem cells, inoculate the embryonic stem cells to the three dimensional model in different cell densities, and observe the phenomena of the forming of embryoid body by self-differentiation of the embryonic stem cells and the law of the embryoid body being differentiated into cells of different ancestries; and the microenvironment in which the embryonic stem cells grow is regulated, the extracellular matrix gel is taken as the main body to carry out the directional differentiation and development research of the embryonic stem cells, thus disclosing the inherent law and mechanism of the in vitro differentiation and development of the embryonic stem cells, and searching for new methods for directional differentiation of embryonic stem cells to the functional cells of important vital organs at the terminal differentiation period; and the invention will provide a new technical method for exploring the development mechanism of the embryonic stem cells.