A method of generating protein-induced pluripotent stem cells by delivering bacterially expressed reprogramming proteins into nuclei of starting somatic cells using the QQ-protein transduction technique, repeating several cell reprogramming cycles for creating reprogrammed protein-induced pluripotent stem cells, moving the reprogrammed cells into a feeder-free medium for expansion, and expanding and passaging the reprogrammed cells in a whole dish for generating homogeneous piPS cells. Also provided are the piPCS cells formed using this method and uses thereof.

The present invention relates to a method for producing cholinergic neurons comprising obtaining neural progenitor cells from stem cells so as to continuously produce cholinergic neural cells with high purity and the same traits, followed by differentiating the neural progenitor cells into the cholinergic neurons, and cholinergic neurons produced therefrom. Since the method of preparing the cholinergic neurons provided in the present invention enables not only production of the cholinergic neurons with high purity, but also rapid production of the cholinergic neurons with the same traits, it can be widely used for effectively treating degenerative cranial nerve diseases such as Alzheimer's disease.

The invention provides in certain embodiments, a method of generating a re-programmed differentiated epithelial cell comprising (a) contacting a non-stem somatic cell obtained from a subject with an effective amount of a de-differentiation agent to form a de-differentiated cell, and (b) transfecting the de-differentiated cell with an expression cassette comprising a promoter operably linked to a nucleic acid encoding a conversion agent to form a re-programmed differentiated cell. The invention also provides in certain embodiments, a method of generating a re-programmed differentiated epithelial cell comprising (a) contacting a non-stem somatic cell obtained from a subject with an effective amount of a de-differentiation agent to form a de-differentiated cell, and (b) contacting the de-differentiated cell with a conversion agent to form a re-programmed differentiated cell. The invention provides in certain embodiments, re-programmed differentiated epithelial cells, and methods of using these re-programmed differentiated epithelial cells to repair or re-generate tissue in vivo.

There are provided to a Rag-2 (Recombination activating gene 2) gene targeting vector, a method for producing SCID-like miniature pigs introduced with the vector, and a use thereof.

A selection method of an iPS cell includes: at a reprogramming process to culture a cell including a plurality of combinations of initializing factors labelled with luminescent genes that are different with each other, acquiring a photon number per unit area or a photon number per unit time of each of the luminescent genes of the cell; judging whether the acquired photon number is more than a threshold that is predetermined for the acquired photon number; and when the acquired photon number is more than the threshold, selecting this cell as an objective cell for a next process.

A method of producing induced pluripotent stem cells including a step for introducing an initialization factor into somatic cells of a mammal, and culturing in a neural stem cell culture medium to obtain induced neural stem cell-like cells, and a step for cultivating said induced neural stem cell-like cells in a growth medium to obtain induced pluripotent stem cells, wherein the initialization factor contains an OCT family, a SOX family, a KLF family, a MYC family, a LIN28 family and a P53 function inhibitor.

The slow kinetics and low efficiency of reprogramming methods to generate human induced pluripotent stem cells (iPSCs) impose major limitations on their utility in biomedical applications. Here we describe a chemical approach that dramatically improves (>200 fold) the efficiency of iPSC generation from human fibroblasts, within seven days of treatment. This will provide a basis for developing safer, more efficient, non-viral methods for reprogramming human somatic cells.

The present disclosure provides compositions and methods of using gene therapy to create induced pluripotent stem cells (iPSCs) without inducing cancer or tumorigenesis. The methods disclosed herein employ plasmids and vectors that contain transcription factors and an anti-oncogene such as PCBP1 which inhibits the expression of cancer biomarkers and concomitant oncogenesis.

In related-art methods of differentiating pluripotent stem cells into a desired cell type, there has not been established a differentiation induction method using human ES/iPS cells and being stable and highly efficient. The use of complicated culture steps is a large problem. In addition, there are also large problems in, for example, that the speed of cell differentiation is low, and hence long-period culture is required, and that the differentiation efficiency is low, and hence it is difficult to obtain a sufficient number of required cells. A method of inducing differentiation into a desired cell type, which induces differentiation within a short period of time and with high efficiency by the use of a Sendai virus vector capable of expressing a transcription factor, and as required, the use of a pluripotent stem cell in which an expression amount of a POU5F1 protein has been substantially removed or reduced, is provided.

Provided here in are methods of producing induced pluripotent stem cells (iPSCs) and isolated population of produced induced pluripotent stem cells (iPSCs). Also provided herein are methods of treating a subject in need thereof using the produced iPSCs or pharmaceutical compositions comprising the produced iPSCs.