122 results about "KLF4" patented technology

Provided herein are methods for generating human induced pluripotent stem cells free from genomic integration of exogenous transgenes by transfecting into nucleated blood cells one or more DNA expression vectors (e.g., plasmid vectors) that do not contain a mammalian origin of replication, and encode and permit expression of one or more reprogramming factors (e.g., Oct4, Sox2, Klf4, and c-Myc). Also provided herein are the integration-free human induced pluripotent stem cells obtained by the methods described herein.

The invention provides compositions and methods useful for deriving or culturing vertebrate ES cells. Certain inventive methods comprise deriving or culturing vertebrate ES cells using medium that comprises a compound that replaces Klf4 or c-Myc in generating iPS cells. The invention provides NOD ES cells and methods of deriving or culturing them.

The invention provides a method for efficiently inducing multipotential stem cells, six inducible factors such as Oct4 (POU5f1), Sox2, c-Myc, Klf4, UFT1, Rex1 (ZFP42), and the like and p53 gene inhibitor are induced into the cells by utilizing a slow virus carrier, thereby the method can efficiently induce and produce the multipotential stem cells.

The invention discloses a novel ips cell inducing method and provides application of inducing genes Oct3/4, Klf4, Sox2, P53, L-Myc, Lin28, RARg and Lrh-1 in inducing isolated mammal somatic cells to generate iPS multipotent stem cells. By an iPS cell establishing method, a method for inducing human iPS stem cells free of exogenous serum, trophoblast cells and animal source is established on the basis of no insertion. The method shortens reprogramming time of the iPS multipotent stem cells and improves cloning generation efficiency, and obtained iPS cells are higher in dryness.

Animal cells, notably adult fibroblasts, are advantageously reprogrammed in direct lineage reprogramming methods using defined factors to produce proliferative and multipotent induced cardiac progenitor cells (iCPC). The iCPC thus produced can be differentiated under suitable differentiation conditions to cardiac lineage cells including cardiomyocytes, smooth muscle cells, and endothelial cells, as evidenced by expression of lineage specific markers. Sets of factors effective in combination to reprogram the fibroblasts can include a set that includes some or all of 5 factors (Mesp1, Baf60c, Nkx2.5, Gata4, Tbx5), a set that includes some or all of 11 factors (Mesp1, Mesp2, Gata4, Gata6, Baf60c, SRF, Isl1, Nkx2.5, Irx4, Tbx5, Tbx20), a set that includes some or all of 18 factors (T, Mesp1, Mesp2, Tbx5, Tbx20, Isl1, Gata4, Gata6, Irx4, Nkx2.5, Hand1, Hand2, Tbx20, Tbx18, Tip60, Baf60c, SRF, Hey2), and a set that includes some or all of 22 factors (T, Mesp1, Mesp2, Tbx5, Tbx20, Isl1, Gata4, Gata6, Irx4, Nkx2.5, Hand1, Hand2, Tbx20, Tbx18, Tip60, Baf60c, SRF, Hey2, Oct4, Klf4, Sox2, L-myc).

The invention discloses a culture medium for establishing a pig iPS cell line and a culture method thereof. A typical pig iPS cell is obtained in the ninth day through transfection by four transcription factors of OCT4, SOX2, KLF4 and c-MYC and induction culture of the culture medium, and the pig iPS cell can be obtained efficiently. The obtained pig iPS cell clone is flat clone, has a regular edge and is similar to the ES cellular morphology of the human body. According to the pig iPS cell line, the pig iPS cell through subculture keeps the undifferentiated state, shows positive in the AP dyeing displaying result and has a pluripotency mark, and the differentiated cell in vitro expresses NCSTN (entoderm), NESTIN (ectoderm) and DESMIN (mesoblast).

The invention discloses a method for preparing pig neural stem cells by reprogramming. The invention provides a method for preparing mammal neural stem cells by reprogramming. The method comprises the following steps: (1) commonly conducting gene OCT3/4, SOX2, KLF4 (Kruppel-Like Factor 4), LMyc and LIN28 into mammal somatic cells by using a non-integrated type episomal carrier to obtain transgenic cells; and (2) culturing the transgenic cells by using an induction culture system containing sodium butyrate to obtain the neural stem cells. An experiment shows that the non-integrated type episomal carrier is used for conducting factors including the OCT3/4, the SOX2, the KLF4, the LMyc, the LIN28 and the like into pig somatic cells; the somatic cells are transdifferentiated into the neural stem cells through optimizing a cultivation condition; the neural stem cells are further differentiated to obtain neural cells with physiological activity so as to lay foundation to nervous system diseases taking a pig as a model.

The present invention concerns the delivery of certain reprogramming factor proteins into cells, such as differenti-atedsomatic cells, in order to induce the epi-genetic reprogramming of the cell so it becomes a pluripotent stem cell. The reprogramming factor protein(s) may be Sox2, Klf4, Oct3/4, c-Myc, Lin28, Nanog, or any protein with reprogramming (-enhancing) activity. These proteins may be linked recombinantly or chemically to a cell penetrating peptide that helps facilitate the introduction of these proteins into the target cell and may be preferably expressed in mammalian cells to maintain them in active forms. Accordingly, the present method of inducing pluripotent stem cell (iPS) formation avoids the use of viral or DNA-based expression vectors or the expression of reprogramming factor genes within target cells, which are known to be harmful to the host target cell and cause cancer.

The invention relates to a hoofed mammal inducible multipotential stem cell and a preparation method thereof. The method comprises the following steps of: A) constructing an expression vector carrying a transcription factor, wherein the transcription factor is Oct4, Sox2, c-Myc, Klf4, Lin28 and Nanog; and B) introducing the transcription factor in the step A) into the cells of the hoofed mammal in a combining form; picking clones of which the form is similar to that of the embryonic stem cell for subculturing; and screening the cell clones meeting the characteristic of the embryonic stem cell to obtain the hoofed mammal iPS cell. The method contributes to determining the most proper culturing condition and the most proper culturing method for establishing an ES cell line of pig, sheep, cattle and other hoofed mammals; and experimental models for various genetic diseases of human beings can be established with the hoofed mammal iPS cell line.

The invention relates to a method for reprogramming cells from aged donors or senescent cells to pluripotent cells that have lost marks of senescence. In particular, the invention relates to an ex vivo method for preparing induced pluripotent stem cells (iPSCs) from a target cell population comprising cells from aged donors or senescent cells, said method comprising the steps of culturing said target cell population under appropriate conditions for reprogramming said cells into iPSCs, wherein said appropriate conditions comprises increasing expression in said target cells, of at least the following reprogramming factors: Oct4, Klf4, Sox2, c-Myc, Lin28 and, optionally Nanog.

Provided are a B cell-derived iPS cell generated using a convenient technique, a technology for providing a human antibody at low cost using the iPS cell, an immunologically humanized mouse prepared using cells differentiated from the iPS cell, and the like. Also provided are a cloned cell obtained by contacting a B cell with nuclear reprogramming factors excluding C/EBPα and Pax5 expression inhibiting substances, particularly nucleic acids that encode Oct3/4, Sox2, Klf4 and c-Myc, wherein the cloned cell has an immunoglobulin gene rearranged therein and possesses pluripotency and replication competence (B-iPS cell). Still also provided are a method of producing a monoclonal antibody against a specified antigen, comprising recovering an antibody from a culture of B cells obtained by differentiating a B-iPS cell derived from a B cell immunized with the specified antigen, and a method of generating an immunologically humanized mouse, comprising transplanting to an immunodeficient mouse a human immunohematological system cell obtained by differentiating a B-iPS cell.

The present invention relates to the production of avian induced pluripotent stem cells from non-pluripotent somatic cells, including embryonic fibroblasts and adult somatic cells. In this method, avian (including quail or chicken) somatic cells are reprogrammed into a state closely resembling embryonic stem cells including the expression of key stem cell markers alkaline phosphatase, etc. by transfecting/transducing the non-stem cells with genes (preferably using a non-integrating vector as otherwise described herein or alternatively an integrating vector, such a lentiviral vector, retroviral vector or inducible lentiviral vector, among others) which express at least nanog, Lin28 and cMyc. In preferred aspects of the invention, the transfected/transduced vectors express nanog, Lin28, cMyc, Oct 4 (POU5F1 or PouV), SOX2 and KLF4. The induced stem cells which are produced contribute to all 3 germ layers, the trophectoderm and in certain aspects, the gonad in chimeric offspring.

2-indolyl imidazo[4,5-d]phenanthroline compounds of Formula I that are capable of intracellular chelation of transition metals and of exerting antiproliferative effects in cancer cells, that are cytostatic and/or cytotoxic, are provided. Compounds of Formula I can also induce apoptosis in cancer cells and are thus capable of exerting a cytotoxic effect on cancer cells. The compounds of Formula I are also capable of selectively inhibiting the proliferation of one or more of prostate cancer cells, colon cancer cells, non-small lung cancer cells and leukemia cells. The compounds of Formula I are also capable of increasing the expression of the zinc-regulated tumour suppressor, KLF4 and thus are useful in inhibiting the proliferation of cancer cells in which KLF4 functions as a tumour-suppressor, including, but not limited to, bladder cancer, cancers of the gastrointestinal tract and various leukemias.

The invention relates to a preparation method of an inducible pluripotent stem cell of a goat. The preparation method comprises the following steps: A) constructing a lentiviral vector carrying a transcription factor which is selected from Oct4, Sox2, c-Myc, Klf4, Lin28 and Nanog; and B) infecting a goat adult cell by combining the transcription factor with the lentiviral vector prepared in the step A, selecting clone with a shape similar to an embryonic stem cell for subculturing, and preparing the inducible pluripotent stem cell of the goat by screening cell clone in accordance with the characteristics of the embryonic stem cell. According to the preparation method, an optimal culture condition and method of a goat ES (embryonic stem) cell line construction can be established; the inducible pluripotent stem cell of the goat is an excellent vector for goat gene targeting, and the inducible pluripotent stem cell of the goat facilitates the revelation of each gene function and the complex development of the goat.

A method is disclosed for the determination of 5-ASA efficacy in preventing and/or treating CRC in a mammalian, which comprises the analysis of the inhibition of the β-catenin pathway in presence of 5-ASA. More in details, the method comprises measuring the expression of at least one gene involved in the regulation of the β-catenin signalling pathway, such as μ-protocadherin, E-cadherin, β-catenin, Axin1, ICAT, p21^waf-1 and the expression of onco-suppressor genes, such as KLF4 and CEBPα. Gene expression can be measured in accordance to the methods commonly available in the art such as QRT-PCR and immunohistochemistry.

Expression of reprogramming factors such as Sox2, klf4, c-myc, Nanog, LIN28 and Oct4 followed by culture in a MEK inhibitor and a GSK3 inhibitor reprograms tissue cells. The invention provides new uses of these inhibitors, for example in inducing completion of the transcriptional resetting of so-called pre-pluripotent (pre-iPS) stem cells, for example as obtained from mammalian neural stem cells or epiblast stem cells treated with single or combinations of the reprogramming factors, expressed transiently or by integrative vectors. Also provided are systems for reprogramming an epiplast stem cells independently of the use of there inhibitors.

The invention relates to a method for improving the inductive generation efficiency of induced pluripotent stem cells, and especially relates to a method for improving the inductive generation efficiency of the induced pluripotent stem cells through using modified histone gene jhdm1a. Jhdm1a and a stem cell induction factor are utilized to improve the induction efficiency and the induction quality of the induced pluripotent stem cells. The stem cell induction factor can be an Oct4 and Klf4 combination, or an Sox2, Oct4 and Klf4 combination, or an Oct4 and Sox2 combination, or individual Oct4. The method also comprises a step that the cells are exposed to vitamin C, so the induction efficiency of the induced pluripotent stem cells is higher than that of methods which do not use vitamin C. The method uses less stem cell induction factor, so the latent carcinogenicity is reduced, a high induction efficiency is obtained, and the induced pluripotent stem cells having a reproduction system transmission capability and a high quality can also be obtained.

The present invention discloses a method for preparing induced pluripotent stem cells by using a Xist Tale inhibitory transcription factor. Synergism of the transcriptional activator-like effectors (Tale)-based inhibitory transcription factor combined to a first intron region of Xist, Oct4, Sox2, Klf4 and c-Myc enhances the efficiency of induction of mouse fibroblasts to the induced pluripotent stem cells. Obtained iPSCs have typical pluripotent stem cell characteristics including involvement in the development of chimeric mice. The method actively promotes the clinical application and researches of the iPSCs, and provides new technical and theoretical bases for studying how to obtain and improve the iPSCs in people, pigs, cattle and other mammals.

The invention provides a preparation method and application of induction type neural stem cells. The preparation method of the induction type neural stem cells comprises the steps of: carrying out overexpression of exogenous factors Ascl1, Neurog2, Pax6, Hes1, Id1, Brn2, c-Myc and Klf4 in sustentacular cells of testis, and culturing in a base culture fluid containing cell growth factors to obtain the induction type neural stem cells. According to the preparation method of the induction type neural stem cells, sustentacular cells of testis derived from the mesoblast are reprogrammed to be neural stem cells derived from the ectoderm, the neural stem cells obtained in the reprogramming manner are proved to have physiological functions in vivo or in vitro, and the cells are safer than pluripotent stem cells for transplantation.

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.

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 present application provides a method for predicting clinical prognosis for a human subject diagnosed with prostate cancer, comprising: detecting an expression level of a marker gene selected from a group consisting of ABCG1, PDCD4, KLF6, ST6, BTD, BANF1, IRS1, ZNF185, ANXA11, DUSP2, KLF4 and DSC2, in a biological sample containing prostate cancer cells obtained from the human subject; and predicting a likehood of the clinical prognosis by comparing the expression level of the marker gene with a reference level. The present application also provides a combination of molecular markers and a kit containing thereof.

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.