49results about How to "Maintain pluripotency" patented technology

The present invention features novel methods for the cryopreservation of mammalian cell that combine the advantages of the slow-freezing and vitrification approaches while avoiding their shortcomings. Generally, the methods include the use of a capillary tube made of a thermally conductive wall material and a thin wall such that the ratio of the thermal conductivity of the wall material to the wall thickness is at least 1,000-500,000. The solution is then exposed to temperatures equal to or less than −80° C. and the vitrification solution containing the mammalian cells is cooled at a rate equal to or greater than 30,000-100,000,000° C. / minute. The exposure of the capillary tube with a thermally conductive and thin wall allows for vitrification of the solution in the absence of ice formation. Cryoprotectants can also be added to the vitrification solution to further prevent ice formation.

Provided are methods and compositions for obtaining genome-engineered iPSCs, and derivative cells with stable and functional genome editing at selected sites. Also provided are cell populations or clonal cell lines derived from genome-engineered iPSCs, which comprise targeted integration of one or more exogenous polynucleotides, and / or in / dels in one or more selected endogenous genes.

[Problem] To provide a cell culture substrate, and a cell culturing method using the substrate and a method for inducing differentiation of pluripotent stem cells using the substrate, which allow culturing of pluripotent stem cells and allow differentiation of pluripotent stem cells into a specified cell species, particularly neural and neural progenitor cells, at a high purity.[Means for Solution] A cell culture substrate, characterized in that, onto the surface, one or more selected from the group consisting of N-cadherin, a fusion protein comprising an entire or partial region of N-cadherin, and a fusion protein comprising an entire or partial region of a protein homologous to N-cadherin are immobilized or coated.

The invention discloses a frozen stock solution of neural stem cells and an application method of the frozen stock solution. The frozen stock solution contains B-27 Supplement without retinoic acid, L-Glutamine, EGF, FGF and vitamin E in Neurobasal-A medium. The frozen stock solution is low in cost; the problem of low thawing rate after existing neural stem cells are frozen is critically solved; and the frozen thawing rate of the neural stem cells is improved.

Provided are methods and compositions for obtaining genome-engineered iPSCs, and derivative cells with stable and functional genome editing at selected sites. Also provided are cell populations or clonal cell lines derived from genome-engineered iPSCs, which comprise targeted integration of one or more exogenous polynucleotides, and / or in / dels in one or more selected endogenous genes.

Provided are methods and compositions for obtaining genome-engineered iPSCs, and derivative cells with stable and functional genome editing at selected sites. Also provided are cell populations or clonal cell lines derived from genome-engineered iPSCs, which comprise targeted integration of one or more exogenous polynucleotides, and / or in / dels in one or more selected endogenous genes.

Described herein are methods and compositions for selection of pluripotent stem cells from a population of mammalian cells (e.g., human) undergoing a process that induces some cells within the population to become pluripotent.

The invention relates to a novel method for culturing a multipotential stem cell without serum and a feeder layer. The method is stable, reliable, and can culture the multipotential stem cell for a long time and also can maintain the self-renewing of the multipotential stem cell. The method is characterized in that a culture system with clear chemical components and without serum and the feeder layer is adopted to perform suspension culture on the multipotential stem cell. According to the morphology of cultured stem cells after 15 generations, mRNA (Messenger Ribonucleic Acid) and protein expression of multipotential marker molecule, and analysis on cell chromatin karyotype, such novel generation system can maintain the properties of the multipotential stem cell during long-term culture. In addition, the differentiative potential of the multipotentail stem cell is further verified by 15-generation suspension culture in an in-vitro embryoid body formation experiment and an in-vivo teratoma formation experiment.

The invention provides a method of effecting de-differentiation of an at least partially differentiated cell or of maintaining pluripotency and / or self-renewing characteristics of an undifferentiated cell. The method comprises increasing the amount or the activity of an Err protein, or a functional fragment thereof, in the cell.

The invention discloses a multi-potent stem cell culture medium. The multi-potent stem cell culture medium comprises a basic culture medium and following added components: L-ascorbic acid-2-magnesium phosphate, sodium selenite, human insulin, ferric citrate, an alkaline basic fibroblast growth factor and a transforming growth factor beta1. The sodium selenite is successfully introduced into a multi-potent stem cell culture medium system to replace human apo-transferrin, so that the production cost of the multi-potent stem cell culture medium can be reduced; exogenous pathogenic risks are reduced; the multi-potent stem cell culture medium can guarantee normal proliferation of cells in a multi-potent stem cell culture process and the multi-potent property of the stem cells can also be maintained.

The invention relates to a method of inducing pluripotency in a responsive mammalian cell, which comprises introducing into the cell an effective amount for initiating pluripotency within the cell of Oct4 protein or a functionally equivalent analogue, variant or fragment thereof. The invention also relates to a method of treatment and / or prophylaxis of a degenerative disease or injury in a mammal, which comprises removing from the mammal one or more responsive cells and culturing the cells in a suitable medium, introducing into the cells an effective amount of Oct4 protein or a functionally equivalent analogue, variant or fragment thereof and subsequently returning the cells to the patient. A further aspect of the invention relates to a method of treatment and / or prophylaxis of a degenerative disease or injury in a mammal, which comprises introducing into responsive cells of the patient an effective amount of Oct4 protein or a functionally equivalent analogue, variant or fragment thereof.

The present invention provides a method for selecting human induced pluripotent stem (iPS) cells which can be safely used for transplantation. That is, the present invention provides a method for selecting human iPS cells having reduced differentiation resistance, comprising the steps of: (1) inducing differentiation of human iPS cells; (2) detecting remaining undifferentiated cells after the step (1); and (3) selecting human iPS cells whose rate of remaining undifferentiated cells detected in step (2) is equivalent to or not more than that of control cells.

A serum-free culture method for human umbilical cord mesenchymal stem cells (hUC-MSC), said method using a step-by-step method to culture hUC-MSC: first using a TME culture medium for culturing for 3-4 hours to promote hUC-MSC adherence, and then switching to a TMD culture medium for rapid amplification.

The invention provides a serum-free feed layer-free mouse induced pluripotent stem cell (iPSC) induction medium and a culture method using the same. The medium is a serum-free feed layer-free medium,can be used as an induction medium for mouse somatic cell-induced formation of iPSCs, improves iPS cell induction efficiency, shortens the induction time and can be used for culturing mouse iPSCs. Inthe culture system, after 20 passages of mouse iPSCs, the mouse iPSCs retain stem cell pluripotency and the normal cell karyotype. An internal teratoma experiment result shows that after culture for 20 passages, the mouse iPSCs in the culture system retain a trilaminar differentiation potential of PSCs.

Methods for deriving and cultivating human embryonic stem (ES) cells and maintaining their pluripotency in culture is provided by utilizing secreted products obtained from the culture medium of human embryonic germ (EG) cell derivatives, such as embryoid body-derived cells. Substrates include compounds such as collagen I, fibronectin, or superfibronectin, or extracellular matrix, typically human derived.

The present invention relates to a method and substance for maintaining the pluripotency and self-replication ability of a stem cell, such as a hematopoietic stem cell, while keeping it undifferentiated. Specifically, the present invention provides a composition for maintaining the expansion or pluripotency of a stem cell, comprising active STAT5, and a method using the same. STAT5 may be in the form of a protein or a nucleic acid. The composition may contain a cellular physiologically active substance (e.g., SCF, TPO, Flt-3L, etc.). The present invention also relates to a cell, a tissue and an organ prepared from the stem cell.

The invention relates to a feeder-free and serum-free human induced pluripotent stem cell isolation culture method. By the feeder-free and serum-free human induced pluripotent stem cell isolation culture method, feeder cells and serum are not needed, a culture medium has clear components, operation is simple, human iPS cells can be efficiently and stably isolated, self-renewal and pluripotency are maintained, and a good experimental foundation is laid for final establishment of a clinical application-level human iPS cell isolation culture system.

The invention relates to a cryopreservation liquid and a cryopreservation method for adipose tissue-derived mesenchymal stem cells ( ADSC). The cryopreservation liquid is composed of the following materials: 3 to 10 mg / ml of vitamine C, 2 to 8 mg / ml of vitamin E, 2 to 8 mg / ml of lipoic acid, 5 to 20 volume percent of human autoserum, and DMEM-LG culture medium in balancing amount. The cryopreservation liquid for ADSC does not contain animal serum and DMSO, and is composed of human autoserum, lipoic acid, vitamin E, vitamin C and other main materials. A cell cryopreserved with the cryopreservation liquid has the advantages of high palinesthesia rate, no impact on cell growth characteristic, and maintanience of stem cell pluripotency after palinesthesia.

The present invention discloses a stem cell culture medium and its applications as well as a stem cell culture method. The said stem cell culture medium contains no serum. The said stem cell culture medium contains amino acids, vitamins, salts, lipids, cytokines and protein polypeptides. The said stem cell culture medium is suitable for rapid culture of stem cells derived from human and mammalian tissues, including but not limited to, adipose mesenchymal stem cells, bone marrow mesenchymal stem cells and umbilical cord blood stem cells. The said culture medium can increase the proliferation speed of the stem cells 3-5 times, without any affects on their differentiation potentials. Comparing the said stem cell culture medium to a routine culture medium, the said culture medium is not only able to proliferate stem cells derived from different sources more rapidly and achieve more proliferation generations, but also keep their differentiation potentials well.

The invention relates to a novel method for culturing a multipotential stem cell without serum and a feeder layer. The method is stable, reliable, and can culture the multipotential stem cell for a long time and also can maintain the self-renewing of the multipotential stem cell. The method is characterized in that a culture system with clear chemical components and without serum and the feeder layer is adopted to perform suspension culture on the multipotential stem cell. According to the morphology of cultured stem cells after 15 generations, mRNA (Messenger Ribonucleic Acid) and protein expression of multipotential marker molecule, and analysis on cell chromatin karyotype, such novel generation system can maintain the properties of the multipotential stem cell during long-term culture. In addition, the differentiative potential of the multipotentail stem cell is further verified by 15-generation suspension culture in an in-vitro embryoid body formation experiment and an in-vivo teratoma formation experiment.

The invention discloses an application of a piperazine derivative as a p53 molecule activity regulator. Mouse and human cell experimental results show that the piperazine derivative can regulate expression of p53 downstream genes, also can make growth and proliferation of normal cells maintained and make proliferation of cancer cells inhibited, and has wide application prospects in the field of cancer therapy. In addition, through regulation of p53 molecules, the piperazine derivative also can adjust a pluripotency signal network of stem cells, inhibit differentiation of the stem cells or maintain the pluripotency of the stem cells, thereby improving the line establishment efficiency of the pluripotent stem cells.

The invention provides a method of effecting de-differentiation of an at least partially differentiated cell or of maintaining pluripotency and / or self-renewing characteristics of an undifferentiated cell. The method comprises increasing the amount or the activity of an Err protein, or a functional fragment thereof, in the cell.

A method of culturing pluripotent stem cells is provided. The method includes culturing pluripotent stem cells in a pluripotent stem cell culture medium supplemented with an additive, where the additive includes a source of acetate ions, a carboxylic acid, or a physiologically acceptable salt of the carboxylic acid, or a combination of these substances, in an amount effective to maintain the pluripotent stem cells in culture in an undifferentiated pluripotent state. Also included are pluripotent stem cell culture media and methods of making such media.

The present invention provides a method for selecting human induced pluripotent stem (iPS) cells which can be safely used for transplantation. That is, the present invention provides a method for selecting human iPS cells having reduced differentiation resistance, comprising the steps of: (1) inducing differentiation of human iPS cells; (2) detecting remaining undifferentiated cells after the step (1); and (3) selecting human iPS cells whose rate of remaining undifferentiated cells detected in step (2) is equivalent to or not more than that of control cells.

A serum-free culture method for human umbilical cord mesenchymal stem cells (hUC-MSC), said method using a step-by-step method to culture hUC-MSC: first using a TME culture medium for culturing for 3-4 hours to promote hUC-MSC adherence, and then switching to a TMD culture medium for rapid amplification.

The invention discloses a vector capable of promoting the self renewal and proliferation of spermatogonial stem cells of a dairy goat and application of the vector. The vector is an eukaryotic expression vector containing a dairy goat gSirt1 gene, wherein the nucleotide sequence of the dairy goat gSirt1 gene is shown as SEQ.ID.NO.1. According to the vector and the application of the vector, over-expression or super expression in the spermatogonial stem cells is realized by the constructed eukaryotic expression vector, and RT-PCR (Reverse Transcription-Polymerase Chain Reaction), Western blot and cell cycle examination show that the gSirt1 gene is helpful for the self renewal, stemness (pluripotency) maintenance and cell proliferation of the spermatogonial stem cells.

The invention discloses a method for promoting bone mesenchymal stem cells transplanted in the rat brain to be directionally differentiated into neurons. The method specifically comprises the steps of: separating mesenchymal stem cells from medulla ossiums and carrying out amplification and purification; cloning into a cDNA (complementary deoxyribonucleic acid ) sequence coded by a human brain derived neurotrophic facto (hBNDF) protein through PCR, constructing an expression vector carrying a hBNDF coding gene, and finally transplanting the mesenchymal stem cells expressing a hBNDF coding gene into the rat brain through a brain stereotaxic method. Under the induction of the hBNDF, the quantity of the mesenchymal stem cells directionally differentiated into the neurons is remarkably increased. According to the method provided by the invention, the quantity of the mesenchymal stem cells which are transplanted in the brain and directionally differentiated into the neurons can be greatly increased, a novel method is developed for the stem cell treatment of the nervous system disease, and a wide clinical application prospect is achieved.