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The present inventions are directed to compositions and methods regarding the reprogramming of biological samples (such as cells) without introducing exogenous genes to the sample. In particular, the present inventions are directed to transducible materials that are capable of transducing into the nuclei and have enhanced retention in the nuclei of the biological sample. The present inventions also are directed to methods of reprogramming a biological sample or treating diseases using the transducible compositions thereof.

The present invention relates to a method for preparing an induced dopaminergic neuronal progenitor (iDP), and a cell therapeutic agent and a composition for treating or preventing Parkinson's disease, containing the iDP prepared thereby as an active ingredient, the method comprising the steps of: inducing Oct4, Sox2, Klf4 and c-Myc gene expression in adult cells; and directly reprogramming the adult cells into iDPs by treating the cells with sonic hedgehog (SHH) and fibroblast growth factor 8 (FGF8).In addition, the present invention relates to a method for preparing mesencephalic dopaminergic neurons, comprising the steps of: isolating a neural stem cell-like colony (NSC-like colony) by culturing the iDP; and making the isolated colony be a single cell, and culturing the same in a neuronal differentiation medium. Additionally, the present invention relates to a method for treating Parkinson's disease or a method for screening an agent for preventing or treating Parkinson's disease, by using the cell of the present invention. An iDP prepared by the method for preparing an iDP of the present invention has remarkably high efficiency for differentiating into dopaminergic neurons compared with known methods, and since the somatic cells of a patient can be used, there are no side effects such as immunogenicity and no ethical problems, thereby enabling the iDP to be widely used in the relevant fields.

The present invention relates to a composition for maintaining the chromosome stability of pluripotent stem cells, containing a small-molecule compound and, more specifically, to a composition for maintaining the chromosome stability of induced pluripotent stem cells, containing a ROCK inhibitor, a MEK inhibitor, an ALK5 inhibitor, or an antioxidant for inhibiting structural and numerical mutations or DNA damage of the chromosomes during a step of inducing pluripotent stem cells, an inducing process, and then culturing of the induced pluripotent stem cells. The composition for maintaining the chromosome stability of induced pluripotent stem cells, containing a small-molecule compound, according to the present invention, increases the dedifferentiation rate and efficiency when the induced pluripotent stem cells are produced and has excellent effects for inhibiting structural and numeral mutations of chromosomes, and is thus very useful for developing an induced pluripotent stem cell therapeutic agent having excelling chromosome stability.

Provided is a method for culturing totipotential stem cells. The method comprises: using an artificial synthetic polypeptide as an extracellular matrix, spreading it on the surface of a cell culture dish, then inoculating the totipotential stem cells onto the surface of the aforementioned culture dish, and injecting in a feeding layer-free culture medium, and then culturing and passaging the inoculated totipotential stem cells. The method provided can support the proliferation of the totipotent stem cells and maintain the proportion of Oct4+ cells at above 80% all the time. After culturing for at least 40 days, namely, after passaging six times and cell doubling about 40 times, the cells still greatly express various markers of the totipotent stem cells, and after proliferation culturing, the karyotype still remains consistent.

A nuclear reprogramming substance (DLX4 gene, OCT3/4 gene, and SOX2 gene) is introduced into a cell, and an iPS cell is produced.

The objective of the present invention is to provide a low-cost, simple method for preparing cells having pluripotency and extremely low tumorigenic risk. Human mesenchymal stem cells from bone marrow (hMSC-BM), human adipose tissue-derived mesenchymal stem cells (hAT-MSC) (also known as "human adipose-derived stem cells (hADSC)")and other such mammal mesenchymal stem cells are cultured in suspension with 7 types of human adherent mature cell (human hepatocyte cells [hHEP cells], human umbilical vein endothelial cells [HUVEC cells], human dermal microlymphatic endothelial cells [HMVEC cells], human epidermal keratinocyte cells [NHEK cells], human bronchial epithelial cells [NHBE cells], human melanocyte cells [NHEM cells], and human smooth muscle cells [UASMC cells]) and 3 types of human adherent precursor cell (human dermal fibroblast cells [NHDF], human skeletal muscle myoblast cells [HSMM cells], and human osteoblast cells [NHOst cells]) in order to form a mass of cells (spheroid), thereby enabling the preparation of cells having pluripotency and extremely low tumorigenic risk.

Described herein are methods for enhancing the nuclear reprogramming of somatic cells to become induced pluripotent stem cells. In particular, the methods disclosed herein involve the use of damage-associated molecular pattern molecules (DAMP). In certain embodiments the DAMPs are aluminum compositions such as aluminum hydroxide. Such DAMPs have unexpectedly and surprisingly been found to enhance the nuclear reprogramming efficiency of the reprogramming factors commonly used to induce somatic cells to become induced pluripotent stem cells. Accordingly, this disclosure describes methods of nuclear reprogramming as well as cells obtained from such methods along with therapeutic methods for using such cells for the treatment of disease amendable to treatment by stem cell therapy; as well as kits for such uses.

The present invention provides a method for improving the efficiency of inducing a pluripotent stem cell, and a vector and composition used for the method. The inventors succeeded in significantly increasing the efficiency of inducing a pluripotent stem cell by further introducing a vector including a KLF gene and not including an OCT gene and a SOX gene in pluripotent stem cell induction including a step for introducing a vector including a KLF gene, an OCT gene, and a SOX gene in this order. This method is capable of efficiently inducing a pluripotent stem cell under temperature conditions closer to a physiological environment, and has excellent characteristics in that the vector is rapidly eliminated after pluripotent stem cell induction. The present invention makes it possible to induce a pluripotent stem cell with greater efficiency.

The present invention relates to a method to reprogramming pluripotent stem cells (PSCs) by epigenetic conditioning and metabolic reprogramming into p PSCs with highly controllable biological functions, the cells obtained by said method as well as methods of using said cells.

The present invention provides a method for improving the efficiency of establishing induced pluripotent stem cells, the method including inhibiting the functioning of STAT1 in a nuclear reprogramming process of a somatic cell.

The present invention relates to a method of redifferentiating adult cells into induced pluripotent stem cells using an electromagnetic field. According to the present invention, the method of redifferentiation into induced pluripotent stem cells using an electromagnetic field has excellent efficacy for redifferentiating adult cells into induced pluripotent stem cells, and induced pluripotent stem cells can be readily obtained. Also, the induced pluripotent stem cells prepared by the method have a large amount of redifferentiation factor expression, and when the stem cells are grafted to a mouse, pluripotency is exhibited. Thus, the redifferentiated induced pluripotent stem cells can be usefully applied to the development of a cell therapeutic agent and studies in the regenerative medicine field.

The present invention relates to a method for inducing customized pluripotent stem cells, pluripotent stem cells prepared by the method, and a composition comprising the pluripotent stem cells, wherein the customized pluripotent stem cells are induced by reprogramming differentiated adult cells by using shikimic acid, a plant extract containing shikimic acid, or an extract of plant stem cells and redifferentiated plant stem cells (callus). According to the present invention, since an ovum is not used to prepare pluripotent stem cells having the same abilities as embryonic stem cells, ethical problems can be solved. Most of all, by using a plant stem cell extract which is proven to be harmless to humans, pluripotent stem cells having ensured safety can be prepared. Also, the present invention can be used to develop an immunocompatible cell therapeutic agent customized for each subject. In addition, it is expected that the present invention can be very useful in studying a therapeutic method and identifying the cause of a disease by inducing pluripotent stem cells from a subject having the disease. Also, the present invention relates to a composition containing shikimic acid or a plant extract containing shikimic acid.

Compositions and methods are described herein for chemically inducing cells that express a single pluripotency transcription factor to change their differentiation state and become cardiac cells, cardiac progenitor cells, cardiomyocytes, or a combination thereof.

A method for culturing and maintaining a pluripotent stem cell in an undifferentiated state is provided. The method comprises culturing the pluripotent stem cell in a medium comprising an MEK inhibitor, a GSK3 inhibitor, a dual inhibitor of AMPK and/or BMP signaling and LIF. A cell produced by the method, cell culture medium and a kit for performing the method described is also provided.

Methods for treating a subject in need thereof are provided which include administering a pharmaceutical composition comprising a protein transduction reagent-modified reprogramming protein to the subject, wherein the protein transduction reagent is non-covalently bound to the reprogramming protein and wherein the protein transduction reagent comprises a cation reagent and a lipid. According to aspects, such methods provide delivery of protein-transduction reagent- modified reprogramming proteins to cancer cells, such as tumor cells, as well as diseased cells of diseased tissues and provide in vivo conversion of diseased cells into normal cells via protein- induced in situ cell reprogramming without administration of nucleic acids to the subject.

Described herein is an inactivated viral particle comprising one or more transcription factor proteins packaged within the particle. A method for using the particle to make induced pluripotent stem cells is also provided.

The present invention relates to a medium composition containing an Ecklonia cava extract for dediffentiating an induced pluripotent stem cell. Also, the present invention relates to a method for producing induced pluripotent stem cells using the medium composition. When using the medium composition, according to the present invention, induced pluripotent stem cells using mesenchymal stem cells can be produced safely, easily, and efficiently, and the pluripotent stem cells which have been produced can be useful as a cell treatment agent by being capable of being differentiated into a variety of cells.

Provided are: a method for evaluating the genomic instability of pluripotent stem cells efficiently and with a high degree of reliability; a method for removing, from a culture of pluripotent stem cells to be evaluated, pluripotent stem cells which are identified as being genomically instable by said evaluation method; and a synthetic peptide that can be used in these methods.　The methods provided by the present invention include: preparing a culture of target pluripotent stem cells; investigating the level of expression of calreticulin in pluripotent stem cells in said culture; and identifying the genomic stability or genomic instability of said stem cells on the basis of said level of expression of calreticulin.

Disclosed herein are methods of generating induced pluripotent stem cells. The method involves providing a quantity of somatic or non-embryonic cells, contacting the contacting the somatic or non-embryonic cells with a quantity of one or more reprogramming factors and one or more RNA molecules, and culturing the somatic or non-embryonic cells for a period of time sufficient to generate at least one induced pluripotent stem cell. Various reprogramming factors and RNA molecules for use in the methods are disclosed herein. Also disclosed are cell lines and pharmaceutical compositions generated by use of the methods.

Provided are a method for preparing an induced pluripotent stem cell and a composition used in the method. The method comprises: introducing a composition for promoting the formation of an induced pluripotent stem cell into a somatic cell, the composition comprising: (i) a c-Jun antagonist and one group of factors from among the following seven such groups: (1) Sox2, Klf4 and c-Myc, (2) Klf4 and c-Myc, (3) Oct3/4, Klf4 and c-Myc, (4) Sox2, Nanog and Lin28, (5) Oct3/4, Nanog and Lin28, (6) Oct3/4, Klf and Sox2, and (7) Klf4 and Sox2; or (ii) the c-Jun antagonist, Jhdm1b and Id1, and at least one of Glis1, Sall4 or Lrh1; or (iii) the c-Jun antagonist, Jhdm1b and Id1, and at least one of: Oct4, Klf4, Sox2, Lin28, Esrrb, Lef1, Utf1 or miRNA C. The present method allows for successful preparation of induced pluripotent stem cells with no generation of abnormal chromosomes.

The purpose of the present invention is to safely culture pluripotent stem cells that are capable of differentiating diversely without mutual cross-contamination. A main transport passage (31) is extended out from a stem cell adjustment area (20), which is provided with a processing chamber (21) for inducing pluripotent stem cells from somatic cells or egg cells or a processing chamber (21) for receiving and adjusting pluripotent stem cells induced at another facility. At least one branch transport passage (32) is branched off of the main transport passage (31) and a cell culturing area (40) comprising stem cell culturing chambers (41-44) and a cultured cell analysis chamber (45) therefor is disposed on each of the branch transport passages (32). Preferably, respective independent worker entrances (22, 47+48, 33) are provided for the stem cell adjustment area (20), the cell culturing area (40), and the transport area (30), and mutual entry and exit of workers between the areas (20, 40, 30) is prohibited.

Provided is a pharmaceutical composition for the treatment of disorders such as Niemann-Pick disease and GM1 gangliosidosis which are caused by the storage of cholesterol, such as lysosomal storage disease. Also provided is a method for screening for said pharmaceutical compositions that uses iPS cell strains that phenocopy phentotypes of these disorders.　Provided is a pharmaceutical composition for the treatment and/or prevention of lysosomal storage disease, characterized by containing hydroxypropyl-γ-cyclodextrin as an active ingredient. Also provided are an iPS cell strain derived from patients suffering from intractable disorders and prepared using a new temperature-sensitive Sendai virus vector, and a screening method for pharmaceuticals using said iPS cell strain.

An antibody that binds a glycosylated protein is disclosed, wherein the glycosylation comprises the glycan motif Fucα1-2Galβ1-3GlcNAc1-3Galβ1 or Fucα1-2Galβ1-3GlcNAc. Antibodies that are cytotoxic against undifferentiated pluripotent cells are also disclosed.

We describe an ELABELA polypeptide comprising a sequence CXXXRCXXXHSRVPFP (SEQ ID NO: 1), in which X signifies an amino acid residue, such as a sequence selected from the group consisting of: SEQ ID NO: 2 to SEQ ID NO: 18, preferably CLQRRCMPLHSRVPFP (SEQ ID NO: 2), or a fragment, homoiogue, variant or derivative thereof, which polypeptide is capable of maintaining self-renewal and/or pluripotency of a stem cell.

The present invention relates to a composition containing Sagunjatang as an active ingredient for promoting the reprogramming of differentiated cells into induced pluripotent stem cells (iPSc), and to a method for preparing the induced pluripotent stem cells using the same. According to the present invention, the Sagunjatang of the present invention effectively promotes the efficiency of induced pluripotent stem cells, and thus can be effectively used in the field of preparing induced pluripotent stem cells.

The present invention relates to an induced pluripotent stem cell (iPS) model for cardiofaciocutaneous (CFC) syndrome, a method for producing the model, and uses in the analysis of neural development in CFC syndrome using the iPS model. Specifically, the CFC syndrome-derived iPS and generation and differentiation of an embryoid body were induced from the fibroblasts of a CFC syndrome patient, and the CFC syndrome-derived iPS and embryoid body were confirmed to exhibit broken embryoid body shapes and no differentiation into neurons, and when a CFC syndrome-derived embryoid body was induced by treating with p-ERK and p-SMAD1 inhibitors, the embryoid body exhibited a normal embryoid body shape and effectively differentiated into neurons, thus CFC syndrome patient-derived stem cell model can be effectively used in the research for neural development in cardiofaciocutaneous syndrome.

The problem is to produce iPS cells under serum-free culture conditions without using feeder cells, and to provide a culture method capable of maintaining the undifferentiated state and pluripotent differentiation ability of iPS cells and other such stem cells over an extended period of time under serum-free culture conditions without using feeder cells. The present invention provides: a method for producing induced pluripotent stem cells (iPS cells), characterized in that iPS cells are induced by culturing reprogrammed somatic cells in serum-free medium without using feeder cells, preferably using fibronectin as an adhesion factor; and a method for maintaining the undifferentiated state and pluripotent differentiation ability of stem cells, characterized in that a protein belonging to the tumor growth factor-β (TGF-β) family is added and stem cells are subcultured in serum-free medium without using feeder cells.

The present invention relates to an induced pluripotent stem cell (iPSC) model for Fabry disease, a preparation method therefor, and a use using the iPSC model in the study of the pathogenesis of Fabry disease and in a method for screening for a Fabry disease therapeutic agent. Specifically, by inducing the creation and differentiation of Fabry disease-derived iPSCs, embryoid bodies (EBs), and vascular cells from fibroblasts of a patient with Fabry disease and confirming the accumulation of globotriaosylceramide (Gb3, CD77) since the expression and activity of GLA protein are significantly reduced in the Fabry disease-derived iPSCs compared with normal cells, and when significantly expressing a marker protein by the differentiation of the Fabry disease-derived iPSCs into vascular endothelial cells and vascular smooth muscle cells as a result of the induced differentiation from the Fabry disease-derived iPSCs into the vascular cells and treating the vascular endothelial cells and the vascular smooth muscle cells with fabrazyme, thereby significantly reducing the accumulation of Gb3, the iPSC model can be usefully used in analytical research for the pathogenic mechanism of Fabry disease and in analytical research for a method for screening for a Fabry disease therapeutic agent.

The invention relates to a method for producing induced pluripotent stem cells (iPS) by culturing somatic cells subjected to a cellular reprogramming method, characterised in that the somatic cells are cultured in the presence of netrin-1 or an analogue of netrin-1 at least at the beginning of the cellular reprogramming method.

Provided is a protein coded by a gene related to cell totipotency and a transcriptional activation domain of a mammalian YAP protein or a fusion protein of a segment with a transcriptional control activity, a coding nucleotide sequence, an expression vector and a composition thereof, as well as a method for inducing the pluripotent stem cells by using the fusion protein.