49results about How to "Maintain pluripotency" patented technology

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.

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.

An object of the present invention is to provide a medium for culturing naive pluripotent stem cells which is for efficiently proliferating naive pluripotent stem cells while maintaining the undifferentiated state and the pluripotency (pluripotent capability) of the naive pluripotent stem cells. The medium of the invention contains a ROCK inhibitor at a concentration of 5 μM or less

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.

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 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 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.

Provided is a serum-free culture medium, the ingredients of the culture medium comprising 0.05-0.2 parts by volume of β-mercaptoethanol, 0.5-2 parts by volume of non-essential amino acid aqueous solution, 4-6 parts by volume of human mesenchymal stem cell culture supernatant concentrate, and 90-95 parts by volume of a-MEM / DMEM-F12 and recombinant human alkaline fibroblast growth factor of a final concentration of 5-5 ng / ml. The present culture medium is used for carrying out stem cell culture.