Kyoto University

Kabushiki Kaisha Kobe Seiko Sho

Keisuke Okita

Shinya Yamanaka

Shushi Ikeda

Shinji Sato

Hideo Hata

Takemasa Yamasaki

Kazutoshi Takahashi

Takeshi Yamada

Tomokazu Nakagawa

Mariko Yamada

Masato Nakagawa

Yusaku Takagawa

Junya Naitou

Kenji Murakami

Masahiro Miyagawa

Tsukasa Shirafuji

Mariko Matsuda

Toshio Murakami

The invention provides an in vitro method of improving the efficiency of establishment of induced pluripotent stem (iPS) cells. The method comprises contacting an isolated somatic cell being reprogrammed into an iPS cell with an inhibitor of p53 function. The invention also provides an in vitro method of producing iPS cells. The method comprises bringing (a) nuclear reprogramming substances or nucleic acids encoding the nuclear reprogramming substances and (b) an inhibitor of p53 function into contact with a somatic cell.

Method of making induced pluripotent stem cells using p53 inhibitors

The present invention provides a production method of iPS cell, including a step of introducing the following (1) and (2):

Highly efficient method for establishing induced pluripotent stem cell

A method of producing an induced pluripotent stem cell includes introducing into a somatic cell one or more non-viral expression vectors. The vectors include one or more of an Oct family gene, a Klf family gene, a Sox family gene, a Myc family gene, a Lin family gene, and Nanog gene. The somatic cell is then cultured in a medium that supports pluripotent stem cells. At least a portion of the one or more introduced non-viral expression vectors is not substantially integrated in the chromosome.

Method of making induced pluripotent stem cells

Provided is a method of producing an iPS cell, comprising bringing (a) Oct3/4 or a nucleic acid that encodes the same, (b) Klf4 or a nucleic acid that encodes the same, and (c) Sox2 or a nucleic acid that encodes the same, as well as (d1) L-Myc or a nucleic acid that encodes the same and/or (d2) a functional inhibitor of p53, into contact with a somatic cell. It is preferable that (a) a nucleic acid that encodes Oct3/4, (b) a nucleic acid that encodes Klf4, (c) a nucleic acid that encodes Sox2, (d1) a nucleic acid that encodes L-Myc and (e) a nucleic acid that encodes Lin28 or Lin28b be inserted into an episomal vector having loxP sequences placed in the same orientation on the 5' and 3&#x2032; sides of a vector constituent essential for the replication of the vector, that (d2) a nucleic acid that encodes an shRNA against p53 be inserted into a vector ensuring transient expression (plasmid vector and the like), and that all these nucleic acids be transferred to a somatic cell.

Method of producing induced pluripotent stem cells using inhibitors of P53

The present invention provides a method of improving the efficiency of establishment of induced pluripotent stem (iPS) cells, comprising inhibiting the p53 function in the step of somatic cell nuclear reprogramming. The inhibition of p53 function is achieved by bringing a substance selected from the group consisting of (1) chemical inhibitors of p53, (2) dominant negative mutants of p53 and nucleic acids that encode the same, (3) siRNAs and shRNAs against p53 and DNAs that encode the same, and (4) p53 pathway inhibitors, into contact with a somatic cell, and the like. The present invention also provides an agent for improving the efficiency of establishment of iPS cells, the agent comprising an inhibitor of p53 function, particularly (1) chemical inhibitors of p53, (2) dominant negative mutants of p53 and nucleic acids that encode the same, (3) siRNAs and shRNAs against p53 and DNAs that encode the same, and (4) p53 pathway inhibitors. The present invention further provides a method of producing an iPS cell, comprising bringing a nuclear reprogramming substance and an inhibitor of p53 function into contact with a somatic cell.

Method of efficiently establishing induced pluripotent stem cells

The present invention relates to a nuclear reprogramming factor having an action of reprogramming a differentiated somatic cell to derive an induced pluripotent stem (iPS) cell. The present invention also relates to the aforementioned iPS cells, methods of generating and maintaining iPS cells, and methods of using iPS cells, including screening and testing methods as well as methods of stem cell therapy. The present invention also relates to somatic cells derived by inducing differentiation of the aforementioned iPS cells.

Induced pluripotent stem cells produced with Oct3/4, Klf4 and Sox2

Somatic cell reprogramming by retroviral vectors encoding Oct3/4. Klf4, c-Myc and Sox2

There is provided a nuclear reprogramming factor for a somatic cell, which comprises a gene product of each of the following three kinds of genes: an Oct family gene, a Klf family gene, and a Myc family gene, as a means for inducing reprogramming of a differentiated cell to conveniently and highly reproducibly establish an induced pluripotent stem cell having pluripotency and growth ability similar to those of ES cells without using embryo or ES cell.

Nuclear reprogramming factor

Growing community of inventors

Kyoto, Japan

Keisuke Okita