50results about How to "High reprogramming efficiency" patented technology

Methods and composition of induction of pluripotent stem cells are disclosed. For example, in certain aspects methods for generating essentially vector-free induced pluripotent stem cells with cell signaling regulators are described. Furthermore, certain aspects of the invention provide novel compositions comprising induced pluripotent stem cells essentially free of exogenous retroviral vector elements in the presence of a medium comprising signaling inhibitors. In certain aspects, feeder-free episomal reprogramming methods may be provided.

Methods and composition of induction of pluripotent stem cells are disclosed. For example, in certain aspects methods for generating induced pluripotent stem cells using reporter genes are described. Furthermore, the invention provides novel reprogramming vectors employing reporter genes.

Methods and composition for providing induced pluripotent stem (iPS) cells are provided. For example, in certain aspects methods including reprogramming B lymphocytes transformed by episomal vectors such as Epstein-Barr virus-based vectors are described. Furthermore, the invention provides induced pluripotent stem cells essentially free of exogenous elements and having B cell immunoglobin variable region rearrangement.

Methods and composition for providing induced pluripotent stem (iPS) cells are provided. For example, in certain aspects methods including reprogramming B lymphocytes transformed by episomal vectors such as Epstein-Barr virus-based vectors are described. Furthermore, the invention provides induced pluripotent stem cells essentially free of exogenous elements and having B cell immunoglobin variable region rearrangement.

Methods and composition of induction of pluripotent stem cells are disclosed. For example, in certain aspects methods for generating induced pluripotent stem cells using reporter genes are described. Furthermore, the invention provides novel reprogramming vectors employing reporter genes.

The invention provides a method for reprogramming ITNK cells based on CRISPR / Cas9. The method comprises a step of transferring an RNP compound into activated T cells through electric conversion to obtain the ITNK cells. According to the method, the RNP compound is transferred into T cells through an electric conversion method; the T cells are converted into ITNK cells; the in-vitro reprogrammingefficiency is improved, the obtained ITNK cells have good in-vitro amplification capacity, the cell amplification multiple of the ITNK cells is 26.8-62.2 times on the fourteenth day after electric conversion, and the ITNK cells have a strong in-vitro killing performance. The reprogrammed ITNK cell preparation capability of the method is improved, so that more clinical patients are benefited.

The present invention relates to a method for increasing the efficiency of inducing pluripotent stem cells, and more particularly, to a method for increasing the efficiency of inducing pluripotent stem cells by utilizing genes Jhdm1b and Jhdm1a that modify histone. By utilizing Jhdm1b, Jhdm1a, and a stem cell inducing factor, the present invention increases the efficiency of inducing pluripotent stem cells and increases the quality of induced pluripotent stem cells. The stem cell inducing factor is a combination of Oct4 and Klf4, or a combination of Sox2, Oct4, and Klf4, or a combination of Oct4 and Sox2, and Oct4 alone. The method of the present invention further comprises exposing the cells to vitamin C, which further increases the efficiency of inducing pluripotent stem cells as compared with the case where no vitamin C is used. By using less stem cell reducing factors, the method of the present invention reduces the potential carcinogenicity, obtains a high inducing efficiency, and provides high-quality induced pluripotent stem cells capable of germ-line transmission.

A method for promoting conversion of cells into cardiomyocytic tissue is carried out by contacting fibrotic tissue (e.g., scar tissue) with a microRNA oligonucleotide or combination of microRNA oligonucleotides. The methods lead to direct reprogramming of fibroblasts to cardiomyocytes or cardiomyoblasts.

The invention provides a method for preparing induced pluripotent stem cells through the reprogramming of somatic cells, and the induced pluripotent stem cells obtained by the method. The method includes the following steps: taking factor Oct4 and Nanog as reprogramming inducing factors to induce into the somatic cells so as to perform reprogramming; and then, cultivating the partially or fully reprogrammed somatic cells in a culture medium containing a specific chemical inducer to obtain the induced pluripotent stem cells. Through the combination of the reprogrammed inducing factors existingin different forms and three small molecular compounds as the specific chemical inducer, the reprogramming efficiency of human somatic cells can be significantly enhanced, and the tumorigenicity of the induced pluripotent stem cells can be reduced.

The invention discloses an induced pluripotent stem cell and a preparation method thereof. The preparation method for the induced pluripotent stem cell comprises the following steps: acquiring somatic cells; constructing a Rab32 overexpression vector and providing transcription factor expression vectors including an Oct4 expression vector, a Sox2 expression vector, a c-Myc expression vector and a Klf4 expression vector; packaging the mixed Rab32 overexpression vector and transcription factor expression vectors so as to prepare infection liquid; and infecting the somatic cell with the infection liquid so as to prepare the induced pluripotent stem cell. The preparation method for the induced pluripotent stem cell is simple and highly efficient and improves the reprogramming efficiency of the somatic cells.

A method for promoting the reprogramming of a non-cardiomyocytic cell or tissue into cardiomyocytic cell or tissue comprising is carried out by contacting a non-cardiomyocytic cell or tissue with a modulator of histone methyltransferase activity or expression.

The invention provides a novel body cell reprogramming inducing method, a kit and an application and relates to an application of an ectodermal differentiation and development correlation factor and / or mesendodermal differentiation and development correlation factor as an inducing factor to generation of an induced pluripotent stem cell. According to the invention, the inducing factor is provided for a differentiated stem cell to induce to generate the induced pluripotent stem cell (iPS cell), and the inducing factor comprises the mesendodermal differentiation and development correlation factor, Sox2, Klf4 and any one c-Myc or the ectodermal differentiation and development correlation factor, Oct4, Klf4 and any one c-Myc or the mesendodermal differentiation and development correlation factor, the ectodermal differentiation and development correlation factor, Klf4 and any one c-Myc. The iPS cell obtained by using the novel body cell reprogramming inducing method has characteristics similar to those of a rat embryonic stem cell, namely has self-renewal maintaining capacity and differentiation omnipotency.

The invention discloses an induced pluripotent stem cell and its preparation method and application. The induced pluripotent stem cell is prepared by introducing a transcription factor and an Erbin gene into skin fibroblasts and carrying out cell culture to allow the skin fibroblasts to undergo reprogramming. Experimental results show that the introduction of the Erbin gene and four genes, i.e., OSKM, into the skin fibroblasts together can improve the reprogramming efficiency of human iPS cells, and the obtained iPS cells can maintain pluripotency for a long time during in-vitro subculturing;and the preparation method provided by the invention overcomes the defects of low reprogramming efficiency and proneness o differentiation during passage of traditional iPS reprogramming methods.

Methods and composition of induction of pluripotent stem cells are disclosed. For example, in certain aspects methods for generating essentially vector-free induced pluripotent stem cells with cell signaling regulators are described. Furthermore, certain aspects of the invention provide novel compositions comprising induced pluripotent stem cells essentially free of exogenous retroviral vector elements in the presence of a medium comprising signaling inhibitors. In certain aspects, feeder-free episomal reprogramming methods may be provided.

The invention discloses a method for inducing reprogramming of skin fibroblast and an application thereof. Transcription factors and an ASF1A-FD genes are introduced into the skin fibroblast, whereinthe transcription factors include Oct3 / 4, Sox2, Klf4 and c-Myc. The ASF1A-FD genes include genes for coding No.1 amino acid-No.154 amino acid of ASF1A protein. By introducing= the ASF1A-FD genes and the OSKM transcription factors (Oct3 / 4, Sox2, Klf4 and c-Myc) into the skin fibroblast, reprogramming efficiency can be greatly improved.

The invention discloses a method for enhancing nonconformable gene expression in human cells. The invention discloses an EBNA-D500DNA molecule, the sequence of which is shown as SEQ IN NO.1. The method disclosed by the invention has the advantage of overcoming the defect that the Episona1 plasmid has lower in human cell line without influencing the unconformability and the characteristic of long-time expression. Moreover, the EBNA-D500DNA is capable of remarkably improving the efficiency of having the Episona1 plasmid to carry out somatic cell reprogramming, can be used for producing human induced pluripotent stem cells which are free of exogenous genes and high in safety, and is applicable to regenerative medicine.

The present invention relates to a method for increasing the efficiency of inducing pluripotent stem cells by utilizing genes Jhdm1a that modify histone. By utilizing Jhdm1a, and a stem cell inducing factor, the present invention increases the efficiency of inducing pluripotent stem cells and increases the quality of induced pluripotent stem cells. The stem cell inducing factor is a combination of Oct4 and Klf4, or a combination of Sox2, Oct4, and Klf4, or a combination of Oct4 and Sox2, and Oct4 alone. The method further comprises exposing the cells to vitamin C, which further increases the efficiency of inducing pluripotent stem cells as compared with the case where no vitamin C is used. By using less stem cell reducing factors, the method of the present invention reduces the potential carcinogenicity, obtains a high inducing efficiency, and provides high-quality induced pluripotent stem cells capable of germ-line transmission.

The present invention relates to a method for increasing the efficiency of inducing pluripotent stem cells by utilizing genes Jhdm1a that modify histone. By utilizing Jhdm1a, and a stem cell inducing factor, the present invention increases the efficiency of inducing pluripotent stem cells and increases the quality of induced pluripotent stem cells. The stem cell inducing factor is a combination of Oct4 and Klf4, or a combination of Sox2, Oct4, and Klf4, or a combination of Oct4 and Sox2, and Oct4 alone. The method further comprises exposing the cells to vitamin C, which further increases the efficiency of inducing pluripotent stem cells as compared with the case where no vitamin C is used. By using less stem cell reducing factors, the method of the present invention reduces the potential carcinogenicity, obtains a high inducing efficiency, and provides high-quality induced pluripotent stem cells capable of germ-line transmission.

The invention provides a method for inhibiting PUMA function and improving iPS (induced Pluripotent Stem) cell construction effect. The method comprises the following steps of: (1) preparing fibroblast MEF (Mouse Embryonic Fibroblast) of a PUMA- / - (PUMA gene knocked out) mice or PUMA- / + mice, and performing passage and frozen storage; (2) converting plasmid PMX-Oct 4, PMX-Sox2, PMX-c-Myc and PMX-KLF4 through DH5 alpha escherichia coli strain, and respectively highly and slightly extracting plasma; (3) packing retrovirus, tittering and performing frozen storage; (4) knocking out reprogramming of embryo fibroblasts MEF of the mice or PUMA- / + mice through PUMA gene; and (5) knocking down the programming of human fibroblast of the PUMPA function. The method has the beneficial effects that the PUMA function is inhibited and the iPS cell construction effect is improved during programming somatic cell; the stability of iPS cell genome can be protected at the same time.

The present invention is in the field of stem cells. Particularly, the present invention relates to a reprogramming method for producing induced pluripotent stem cells (iPSC) by transducing somatic differentiated cells with a viral vector expressing Δ133p53β isoform, Δ133p53γ isoform or both Δ133p53β and Δ133p53γ isoforms; to induced pluripotent stem cells as produced by this method; to uses thereof in cell therapy and as an in vitro model for studying various diseases; and to a method for detecting iPSC.

A method for promoting conversion of cells into cardiomyocytic tissue is carried out by contacting fibrotic tissue (e.g., scar tissue) with a microRNA oligonucleotide or combination of microRNA oligonucleotides. The methods lead to direct reprogramming of fibroblasts to cardiomyocytes or cardiomyoblasts.

The invention discloses a somatic cell reprogramming method. The somatic cell reprogramming method comprises the following steps of preparing somatic cells from different sources, so that one or moreof Sox2, Klf4, Oct4 and c-Myc can be expressed; preparing a culture medium for programming: adding serum replacement additives, growth factors and micromolecule compounds to a basic culture medium; then, adding compounds to the basic culture medium, and performing uniform mixing reversely; and culturing somatic cells in the culture medium for programming to obtain induction multipotential stem cells. According to the method, the efficiency that the somatic cells are reprogrammed into the induction multipotential stem cells can be greatly improved. The method is used, so that transition of somatic cell mesenchyma into epithelia in the serum-free culture medium can be realized, the reprogramming efficiency of the somatic cells of different sources can be improved, and a technique support isprovided for application of induction of multipotential stem cells to regeneration medicines in the future.

Methods for producing engineered induced pluripotent stem (iPS) cells are provided comprising introducing a first nucleic acid into somatic cells for integration into their genome and reprogramming the cells to produce engineered iPS cells having the nucleic acid integrated into their genome. For example, in certain aspects the cells are reprogrammed by introduction of a genetic element that expresses one or more reprogramming factor and culturing of the cells under conditions sufficient to produce reprogrammed cells.

The invention provides a culture medium for induction somatic cell reprogramming, the culture medium includes a culture medium of normal induction somatic cell reprogramme and the phosphatidic acid. The invention also provides a method for induction somatic cell reprogramming, the method includes the culture substrate used in any period of the induction somatic cell reprogramme or any period of the induction somatic cell reprogramme, and adds the phosphatidic acid in the culture substrate. The study found the phosphatidic acid as the most simple lipoid, by adding the phosphatidic acid into the culture substrate of the induction somatic cell reprogramme, the method can obviously increase the reprogramming efficiency of somatic cells, and adds to the lowering of cell apoptosis in the reprogramming process, thereby can obtain large quantity of high quality iPS cells, and builds a foundation for application of PS cells in drug screening and in future reform medicine.

A method for promoting the reprogramming of a non-cardiomyocytic cell or tissue into cardiomyocytic cell or tissue comprising is carried out by contacting a non-cardiomyocytic cell or tissue with a modulator of histone methyltransferase activity or expression.

The invention discloses a cell reprogramming method and belongs to the technical field of induced pluripotent stem cells. The method comprises the steps of on-chip cell reprogramming, on-chip clone selection and on-chip microenvironment optimization. The method can obviously improve the cell reprogramming efficiency. Produced clones have higher quality, and the occurrence positions of the clones are controllable and more uniform instead of being random as before, so that realization of subsequent industrialized standards is facilitated. By means of the method, cells can be conveniently tracked, the dynamic change process of reprogramming is monitored, and therefore convenience is provided for understanding the mechanism of reprogramming. The method can achieve precise control over the micro-pore physical and chemical environment at high flux, and screening at the high flux is favorable for the culture micro-environment of reprogramming.

The invention discloses an induced pluripotent stem cell and its preparation method and application. Transcription factors and Erbin gene are introduced into skin fibroblasts, and the skin fibroblasts are reprogrammed through cell culture to obtain induced pluripotent stem cells . The experimental results show that introducing Erbin gene and OSKM four genes into skin fibroblasts can improve the reprogramming efficiency of human iPS cells, and the obtained iPS cells can maintain pluripotency for a long time in in vitro culture and passage, solving the problem of traditional iPS cells. The reprogramming method has the defects of low reprogramming efficiency and easy differentiation during passage.

The invention provides a method for improving the reprogramming efficiency of somatic cells, a method for improving the in vitro development rate of a cloned embryo, a method for improving the maturedevelopmental rate of the cloned embryo, a method for improving the establishment efficiency of in vitro nuclear transplantation of non-human embryonic stem cell line, and an in vitro processing method for a reconstructed embryo used for embryo transplantation, which all adopt m-carboxycinnamic acid bishydroxamide to treat the reconstructed embryo. The invention also provides application of the m-carboxycinnamic acid bishydroxamide in the preparation of a medicament for improving the reprogramming efficiency of the somatic cells.

The invention provides a method for improving the reprogramming efficiency of somatic cells, a method for improving the in vitro development rate of a cloned embryo, a method for improving the mature developmental rate of the cloned embryo, a method for improving the establishment efficiency of in vitro nuclear transplantation of non-human embryonic stem cell line, and an in vitro processing method for a reconstructed embryo used for embryo transplantation, which all adopt m-carboxycinnamic acid bishydroxamide to treat the reconstructed embryo. The invention also provides application of the m-carboxycinnamic acid bishydroxamide in the preparation of a medicament for improving the reprogramming efficiency of the somatic cells.

The invention discloses a method for enhancing nonconformable gene expression in human cells. The invention discloses an EBNA-D500DNA molecule, the sequence of which is shown as SEQ IN NO.1. The method disclosed by the invention has the advantage of overcoming the defect that the Episona1 plasmid has lower in human cell line without influencing the unconformability and the characteristic of long-time expression. Moreover, the EBNA-D500DNA is capable of remarkably improving the efficiency of having the Episona1 plasmid to carry out somatic cell reprogramming, can be used for producing human induced pluripotent stem cells which are free of exogenous genes and high in safety, and is applicable to regenerative medicine.