An objective of the present invention is to provide an improved negative-strand RNA viral vector and a use thereof, the negative-strand RNA viral vector exhibiting transient high expression of genes loaded in the vector and enabling the rapid removal of the vector after said expression. It was discovered that by adding a micro-RNA target sequence to the NP, P, or L gene of a negative-strand RNA viral vector, it is possible to control the expression of the vector depending on the micro-RNA expressed by the introduction cell. In particular, when a micro-RNA target sequence was added to the NP or P gene, the expression of the vector decreased depending on the micro-RNA, and the removal of the vector was promoted, while the effect was reversed when a micro-RNA target sequence was added to the L gene. The vector can be applied in cell therapy and regenerative medicine and can be used as a therapeutic vector that targets cancer.

To provide a method for producing induced pluripotent stem cells (iPS cells) that can initialize somatic cells without using feeder cells or a substrate. For production of iPS cells, the following steps are carried out: I. introducing an initialization gene into a somatic cell; and II. performing initialization and amplification culture of the cell into which the gene has been introduced, in a liquid medium comprising at least one of a protein kinase C (PKC) inhibitor and a WNT inhibitor under a suspension culture condition.

This invention relates to retroviral gene transfer vectors, particularly lentiviral vectors, pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, comprising a promoter and a transgene; and methods of making the same. The present invention also relates to the use of said vectors in gene therapy, particularly for the treatment of respiratory tract diseases such as Cystic Fibrosis (CF).

The invention provides compositions and methods for manufacturing pluripotent cells. In particular, the invention provides improved culture platforms for manufacturing pluripotent cells with ground state pluripotency.

The present invention provides a method for preparing induced pluripotent stem cells through somatic cell reprogramming and induced pluripotent stem cells obtained therefrom. The present method comprises introducing the factors Oct4 and Nanog as reprogramming-inducing factors into somatic cells to perform reprogramming; followed by culturing the partially or fully reprogrammed somatic cells in a medium comprising specific chemical inducing agents to obtain induced pluripotent stem cells. In the present invention, the combination of different forms of reprogramming-inducing factors and three small-molecule compounds as chemical inducing agents can significantly improve the reprogramming efficiency of human somatic cells and reduce the tumorigenicity of the obtained induced pluripotent stem cells.

The present invention relates to: a method for producing immunocytes, specifically induced natural killer T (iNKT) cells that are induced by direct reprogramming of isolated somatic cells, and chimeric antigen receptor (CAR)-iNKT cells into which a CAR gene encoding a CAR is introduced; iNKT cells produced by the method; and a cell therapy composition and a pharmaceutical composition for preventing or treating cancer, comprising the iNKT cells. The method according to the present invention can produce, through direct reprogramming, iNKT cells or iNKT cells into which a CAR gene is introduced, from isolated cells so as to simplify the production process and shorten production time, thereby reducing costs, to have excellent NKT cell production efficiency, and to ensure safety according to the production without passing through induced pluripotent stem cells, thereby having an excellent NKT cell production effect distinguished from that of a conventional reprogramming technique. In addition, the iNKT cells or iNKT cells into which a CAR gene is introduced, which are produced by the method, have an excellent cancer cell killing ability, and thus can be effectively used as a cell therapy composition or a pharmaceutical composition for preventing or treating cancer.

The present invention provides a temperature-sensitive negative-strand RNA virus or virus vector and an RNA genome thereof. According to the invention, provided is a negative-strand RNA virus or virus vector having a negative-strand RNA genome wherein the phosphoprotein (P protein) on the RNA genome has an amino acid mutation(s) corresponding to a substitution(s) in an amino acid(s) of the P protein corresponding to one or more or all of D433, R434, and K437 and optionally further has an amino acid mutation corresponding to a further amino acid substitution in an amino acid of the P protein corresponding to L511.

The present invention addresses the problem of providing a chimeric envelope protein that pseudotypes a virus, and also providing efficient gene transfer and gene expression techniques to lymphocytes such as B cells, CD4 positive T cells, and CD8 positive T cells contained in peripheral blood and immortalized cells derived from these cells, said techniques being characterized by using an RNA virus vector having the aforesaid chimeric protein. In a gene transfer method using a single-stranded RNA virus vector such as a Sendai virus vector or a stealth RNA vector, the virus is pseudotyped by using, as the envelope proteins of viral particles, a chimeric F protein having a morbillivirus-derived F protein region and a chimeric H protein having a morbillivirus-derived H protein region.

Provided is a cell culture method including introducing a factor into cells in a cell culture vessel, and culturing the cells into which the factor has been introduced in the same cell culture vessel. Also provided is a mononuclear cell collection method including treating blood to prepare a treated blood from which erythrocytes have been at least partially removed, diluting the treated blood, causing sedimentation of mononuclear cells contained in the diluted treated blood, removing the supernatant from the diluted treated blood, and collecting the mononuclear cells.

Provided is a production method for induced pluripotent stem cells in which induced pluripotent stem cells having high undifferentiation and differentiation potency are easily obtained. A production method for induced pluripotent stem cells according to the present invention includes the steps of: introducing an reprogramming factor into somatic cells; and culturing the somatic cells into which the reprogramming factor has been introduced in the presence of a cell scaffold containing a peptide-conjugated polyvinyl acetal resin.